upta G

Biofertilizers: A novel tool for agriculture

With the increasing demand in agriculture it has become important for us to increase the productivity by using various fertilizers insecticides Pesticides .But with the tremendous use of these products the soil has been affected badly because of the depletion in the essential minerals of the soil. So to overcome this problem it has become important for all of us touse a different remedy for the production of various biofertilizers. The production if such biofertilizers from various microorganisms was studied from this work. Introduction About 100 years ago, Hellriegal and Wilfarth demonstrated clearly that fixation of atmospheric nitrogen takes place in legumes, although earlier in 1980s, Boussingault, a French agriculturist, provided the data to show that legumes are superior to cereals in furnishing the nitrogen to plant. Beijerinck, a Dutch scientist, in 1888 isolated root nodule bacteria, (Rhizobium) from root nodules of legumes and that the time many other have defined. Bio-fertilizers, in strict sense, are not fertilizers, which directly give nutrition to crop plants. These are cultures of microorganisms like bacteria, fungi, packed in a carrier material. Thus, the critical input in biofertilizer is the microorganisms. They help the plants indirectly through better Nitrogen (N) fixation or improving the nutrient availability in the soil. The term “Biofertilizer” or more appropriately a “Microbial inoculants” can generally be defined as preparation containing live or latent cells of efficient strains of Nitrogen fixing, Phosphate solublising or cellulolytic microorganisms used for application to seeds, soil or composting areas with the objective of increasing the number of such microorganisms and accelerate those microbial process which augment the availability of nutrients that can be easily assimilated by plants. Biofertilizer can provide an economically viable support to small and marginal farmers for realizing the ultimate goal of increasing productivity. Biofertilizer are low cost, effective and renewable source of plant nutrients to supplement chemical fertilizers. Microorganisms, which can be used as biofertilizer, include bacteria, fungi and blue green algae. These organisms are added to the rhizosphere of the plant to enhance their activity in the soil. Sustainable crop production depends much on good soil health. Soil health maintenance warrants optimum combination of organic and inorganic components of the soil. Repeated use of chemical fertilizers destroys soil biota. In nature, there are a number of useful soil microorganisms that can help plants to absorb nutrients. Their utility can be enhanced with human intervention by selecting efficient organisms, culturing them and adding them to soils directly or through seeds. The cultured microorganisms packed in some carrier material for easy application in the field are called biofertilizers. Bio-fertilizers are living microorganisms of bacterial, fungal and algae origin. Their mode of action differs and can be applied alone or in combination. By systematic research, efficient strains are identified to suit to given soil and climatic conditions. Such strains have to be mass multiplied in laboratory and distributed to farmers. They are packed in carrier materials like peat, lignite powder in such a way that they will have sufficient shelf life. The list of commonly produced bio-fertilizers in our country is shown in Table no 1. The biofertilizer are mainly purchased by State Agriculture Departments and distributed to the farmers at concessional rates. About 200 to 500 grams of carrier material is only needed per acre, costing about Rs.10/to 25/-. The current trends indicate that there is a steady increase in the demand in the Southern states except Andhra Pradesh, Western States and Madhya Pradesh and Rajasthan. Decomposing Culture Composting is a process essentially meant to utilize soil waste of animal-plant origin. Composting procedure can be hastened by inoculating the residue with cellulolytic – lignolytic microorganisms such as Trichoderma viridae, Aspergillus niger, Aspergillus terrus etc. The improvement in soil fertility due to sludge Biofertilizers: A novel tool for agriculture International Journal of Microbiology Research, ISSN: 0975-5276, Volume 1, Issue 2, 2009 24 application was reflected through population diversity of soil microbial community. The technologies used were indigenous and Agricultural Universities and Research Laboratories of GOI standardize the scientific aspects of production. Machineries and laboratory equipments are available from various manufacturers and are of BIS standards. The primary objective of biofertilizer projects could be production of various strains of good quality biofertilizer using most modern technology. The infrastructure and laboratory facilities created, however, can be utilized for the production of bio pesticides and bio control agents. Multi product range will increase the viability. In line with the technology and objective of biofertilizer production, various facilities are required for the successful implementation of such projects .From foregoing paragraphs, it is cleared that two organisms, via a nitrogen fixer and a phosphate solubilizers can be mixed and applied. Composite cultures containing both nitrogen-fixer and phosphate solubilizers are an area that holds considerable importance. Attempts were made to prepare a biofertilizer from forest waste using EM solution (effective microorganism). Forest waste like weeds, stalks, stems, fallen leaves, pruning, dead branches and grass clippings, old flowers, green pruning etc. EM solution contains combination of Phototrophic Bacteria, Lactic acid bacteria and Yeast. Photosynthetic bacteria synthesize useful substances from secretions of roots, organic matter and/or harmful gases (e.g. hydrogen sulfide), by using sunlight and the heat of soil as sources of energy. Lactic acid bacteria produce lactic acid from sugars and other carbohydrates, developed by photosynthetic bacteria and yeast. Yeast synthesizes antimicrobial and other useful substances required for plant growth from amino acids and sugars secreted by photosynthetic bacteria, organic matter and plant roots. Materials and Methods Preparation of biofertilizer All organic material wastes available on a forest, such as weeds, stalks, stems, fallen leaves, pruning, and dead branches were collected. Hard woody material such as dead branches and pigeon pea stalks were first crushed with the help of crusher machine before being piled. Material composts best when it is 1.25-3.75 cm in size. Soft, succulent tissues did not need chopping into very small pieces because they decompose rapidly. The harder or woodier the tissues, the smaller they need to be in order to decompose rapidly. Chopping material with a sharp shovel is effective, when pruning plants, the material were cut into small pieces using the pruning shears. This requires a little effort but the results are worth it. For the composting process to work most effectively, the material to be composted should have a C: N ratio of 20:1. Mixing equal volumes of green plant material with equal volumes of naturally dry plant material yields such a ratio. The green material can be grass clippings, old flowers, green pruning, weeds, fresh garbage and fruit and vegetable wastes. The dried material can be fallen leaves, dried grass, and woody material from pruning. All the ingredients are mixed together, except accelerator (EM solution).These grind particle mix with soil (soil used because soil has ability to absorb moisture and provided suitable condition for growth of microorganism) mix the mixture uniformly. 2. Pit site and size: -The site of the compost pit was taken a level high enough to prevent rainwater from entering in the monsoon season; a temporary shed was to be constructed over it to protect the compost from heavy rainfall and to avoid direct contact of environmental heat during summer. The pit was made about 1m deep, 1.5-2 m wide and of a suitable length can vary according to the availability of land. The pit was having a slant walls and floor with 90-cm slope to prevent water logging. 3. Filling the pit:Before making first pile in the pit, base of the pit was sprinkled with water so that it will help in maintain optimum temperature for the growth of microorganism. Adding Organic residues form first pile with soil in pit. A unit pile is about 5m (length) × 1m (width) × 1m (height) in size. The pile is sprinkled with water for adequate moisture content and EM solution (from maple orgtech India ltd, kolkata) is sprinkled on it. This procedure is repeated until the pit is full. The pit is covered with a plastic sheet. To maintain the moisture condition daily adequate amount of water is added around the pit, this continues for two to three weeks. After two or three weeks later, the whole pile was mixed in order to boost aerobic decomposition. 4. The pile was turned: The pile was turned after two weeks and then again after another week. Normally, the compost is ready after two weeks when the heap has cooled down and the height of the pile has fallen to about 70 cm. Methods of Analysis Of Organic Fertilizer 1. Estimation oF pH: 25g compost was mixed with 50ml of distilled water and kept on rotary shaker for 2 hours. Filtrate was obtained through Whatman filter paper under vacuum using a funnel. pH of filtrate is determined using pH meter. 2. Estimation of moisture:5 gm of prepared compost was taken on a dry petri-dish. It was heated in an oven for about 5 hours at 65 C, constant weighing was done. Cooling is done in a desicator and weigh. Percentage loss in weight was estimated as moisture content of the Biofertilizer. Calculation of Moisture:Moisture percent by weight = 100(B-C)/ B-A

Optimization of production and partial purification of laccase by Phanerochaete chrysosporium using submerged fermenation

Laccases are multi copper oxidases having wide substrate specificity mainly found in white-rot fungi, which are the only microorganisms able to degrade the whole wood components. In contrast to most enzymes, which are generally very substrate specific, laccases act on a surprisingly broad range of substrates, including diphenols, polyphenols, different substituted phenols, diamines, aromatic amines, benzenethiols and even some inorganic compounds such as iodine. As they are capable of degrading a wide variety of compounds they are commercially very significant. This project aims at studying the production optimization of laccase using different carbon sources.

Optimization of growth and production of protease by Penicillium species using submerged fermentation

Enzymes play a vital role in the industry with a wide range of application. Agricutural waste is the maximum waste been produced in India. This Project aims at using various agricultural wastes for the production of Protease. To achieve this various research is being conducted. Various Bacterial and Fungal species are being used for the production of Protease. Penicillium species was being used in this work for the production of Protease .In this work the various components of the media were studied and were optimized and used for the production of Protease.

Study of pectinase production in submerged fermentation using different strains of Aspergillus Niger

Fermentation technology plays a very important role in the field of Biotechnology. Production of various enzymes, antibiotics, single cell proteins, various food products, etc is carried out in a vessel called as the bioreactor which is the seed of fermentation technology. The project includes study of Pectinase production using submerged fermentation and various parameters like different substrate, different strains and fermentation time to obtain the maximum yield of Pectinase. To achieve this various research is being conducted. Aspergillus Niger species was being used in this work for the production of Pectinase.

Orange: Research analysis for wine study

Fermentation of citrus juice may offer a relative simple avenue for reducing post harvest wastage of citrus fruits in low utilization environment and in places where the production of citrus concentrates is low or nonexistent. Orange juice concentrates are readily storable and can be used for production processes even when the fruit is out of season and investigated the possibility of exploiting the fermentative ability of yeasts to produce orange wines.

Immunogenomics: recent discoveries

Immunogenomics is a new research field, which addresses the interface between host and (pathogen) proteome, bridging informatics technology, genomics, proteomics, immunology and clinical medicine. The switch from the paradigm of solely hypothesis-driven immunological research to a more interactive and flexible relationship between classical research and a new discovery-driven approach is termed immunogenomics. This review will focus on the immunogenomics and recent discoveries in Immunogenomics, which are leading to insights into novel immunomodulatory therapies. The research regarding the different aspects of immunogenomics; rather than providing a comprehensive review, we will highlight various factors that influence immunogenomics and how modulation of the immune response may be incorporated into future trends. Keywords- Immunogenomics, Genetic regulation, MS, GC, NMR, microarray

Construction and standardization of a bioreactor for the production of alkaline protease from Bacillus licheniformis (NCIM-2044)

Standardization of a Bioreactor for Industrial purpose is of prime importance. The construction of the Bioreactor needs a proper understanding of the different parts of the bioreactor and how they could be used. Hence the basic focus of this work was the Construction and the Standardization of the Bioreactor. The Production of Protease was done by altering the pH and checking the KL a level in the fermentation Process. Introduction Enzymes are biocatalysts, that catalyze (i.e. accelerate) chemical reactions [1]. In enzymatic reactions, the molecules at the beginning of the process are called substrates, and the enzyme converts them into different molecules, the products. Bacillus licheniformis is a Grampositive, spore-forming bacterium widely distributed as a saprophytic organism in the environment. B. licheniformis is a facultative anaerobe, which may allow it to grow in additional ecological niches. Optimal temperature for enzyme secretion is between 37°C to 41°C. Component parts of a fermentation process Regardless of the type of fermentation an established process may be divided into six basic component parts; • The formulation of media to be used in culturing the process organism during the development of the inoculum and in the production fermenter. • The sterilization of the medium, fermenters and ancillary equipments. • The production of an active, pure culture in sufficient quantity to inoculate the production vessel. • The growth of the organism in the production fermenter under optimum conditions for product formation. • The extraction of the product and its purification. • The disposal of effluents produced by the process. Bioreactor: Bioreactor (fermenter) is the heart of the fermentation process. In designing and construction of a bioreactor a number of points must be considered; Construction Materials: In fermentations with strict aseptic requirements, generally, it is important to select materials that can withstand repeated steam sterilization cycles. Two basic types of fermenters are used;va glass vessel with a round or flat bottom and a top flanged carrying plate. All vessels of this type have to sterilized by autoclaving. A glass cylinder with stainless steel top and bottom plates and these fermenters may be sterilized in situ, but 30 cm diameter is the upper size limit to safely withstand working pressures [10]. Temperature Control: Normally, in the design and construction of a fermenter there must be adequate provision for temperature control which will affect the design of the vessel body as heat will be produced by microbial activity and mechanical agitation [10]. Aeration and Agitation The type of aeration-agitation system used in a particular fermenter depends on the characteristics of the fermentation process under construction. The structural components of the fermenter involved in aeration and agitation are: • The agitator (impeller). • The aeration system (sparger). • Baffles. Volumetric Mass-Transfer Coefficient (KLa): The oxygen demand of an industrial fermentation process is normally satisfied by aerating and agitating the fermentation broth.. Rate of oxygen transfer from air bubble to the liquid phase may be described by the equation; dCL/dt = KLa(C* CL) Where, CL is the concentration of dissolved oxygen in the fermentation broth (Mmoles dm-3) t is time (hours) Construction and standardization of a bioreactor for the production of alkaline protease from Bacillus licheniformis (NCIM-2044) International Journal of Microbiology Research, ISSN: 0975-5276, Volume 1, Issue 2, 2009 33 dCL/dt is the change in oxygen concentration over a time period i.e., oxygen transfer rate (Mmoles O2 dm-3h-1) KL mass transfer coefficient (cmh-1) a is the gas/liquid interphase area per liquid volume (cm2 cm-3) C* is the saturated dissolved oxygen concentration (mmoles dm-3) [10] Determination of KLa values: The determination of the KLa of a fermenter is essential in order to establish its aeration efficiency and to quantify the effects of operating variables on the provision of oxygen. The following are the techniques used for determination of KLa; • Sulphite oxidation technique • Gassing-out technique • The Static Method of Gassing-out • The Dynamic Method of Gassing-out The major factors involved in scale-up are: • Inoculum development • Sterilization • Environmental parameters • Nutrient availability, • pH, • Temperature, • Dissolved oxygen concentration, • Shear condition, • Dissolved carbon dioxide concentration • Foam production All the above parameters are affected by aeration and agitation, either in terms of bulk mixing or the provision of oxygen. Points a, b, c and e are related to bulk mixing whilst d, e, f, & g are related to air flow and oxygen transfer. Modes of Bioreactor Operations: Bioreactors can be operated in following three ways; • Batch bioreactors • Continuous bioreactors Fed-batch bioreactors Materials and Method Organism and growth conditions: Stock cultures of Bacillus licheniformis, (NCIM 2044) obtained from N.C.L, Pune were maintained and subcultured weekly on Nutrient Agar slants. Inoculum preparation: A loopful of culture was taken out from the Nutrient Agar slant with the help of a sterile nichrome wire loop under aseptic conditions, and was used to inoculate the Alkaline Protease Producing Broth (APPB) which had already been autoclaved before inoculation. Shake flask studies: A 1.0 ml portion of the above inoculum was transferred to 8 different 250 ml side-arm flasks containing 100 ml of APPB each, with pH range varying from 7.5 to 11.0 respectively under complete aseptic conditions. Two type of centrifugation process were carried out in order to get the supernatant containing the extracellular enzyme; • In first process, the samples were centrifuged at 4000 rpm for 40 mins. • In second process, the samples were centrifuged at 16000 rpm for 40 mins at 4 C. Measurement of proteolytic activity: Proteolytic activity of the organisms, recovered from the 8 different side-arm flasks of various pH ranges was determined on the basis of their ability to produce a clear zone of hydrolysis on skim milk agar plates by agar ditch method. Diameter of the zone of clearance was measured in mm. Estimation of total extracellular soluble protein: The total extracellular soluble protein content present in the supernatant was quantitatively estimated according to the Bradford method of protein estimation [9]. • The above mentioned solutions after preparation were vortexed and mixed thoroughly. • The samples were prepared as 40 μl sample + 760 μl of distilled water + 200 μl Bradford reagent (volume was made up to 5 ml as previously mentioned). • The spectrophotometer was set at a wavelength of 595 nm. • The blanks were first added into clean glass cuvettes and the instrument was auto zeroed. • Firstly, the standard solutions were added into the cuvettes and the O.D. was recorded for all the standard solutions. • Subsequently, the samples were added into the cuvettes and the O.D. of the same was recorded. • A graph of concentration vs. O.D. was plotted and the values of x and y were calculated. • Using the values of x and y the concentration/200 μl was calculated. Quantification of Alkaline Protease: Protease activity was determined spectrophotometrically by the Anson method, with a slight modification. The samples were prepared by mixing 0.1 ml enzyme (supernatant) with 1.0 ml (0.5 % casein) and 1.9 ml 0.1 M Tris-HCl Buffer, at 37°C for 30 min, and then the reaction was arrested by the addition of 2.0 ml of 5% trichloroacetic acid (TCA). This mixture was centrifuged at 3500g for 20min, and the released amino acids (in supernatant) were measured as tyrosine by Folin-Lowry method. The further tests for FolinLowry were carried out as follows; Firstly, a stock solution of BSA was prepared at a concentration of 0.2 mg/ml, which was used for preparing the standard dilutions for the assay. The standards and the samples were prepared according to the table listed below: Substrate utilization: Substrate utilization was carried out in order to determine the uptake of glucose from the fermentation medium by the cells for biomass production. The yield factor (Y)

Biostatistical analysis of the HIV-aids patients with reference to various factors.

The prevalence of HIV /AIDS in Indian population has been carried out. However analysis of secondary infection after HIV is carried out by the present studies. A total of 58 patients suffering from HIVAIDS were analyzed with reference to the age, gender, BMI and Mode of infection. Data analysis showed that patients suffering from HIV / AIDS have greater risks of Tuberculosis disorders. A distinct correlation between Tuberculosis and HIV/ AIDS was observed during the present studies. KeywordsHIV, Tuberculosis Introduction Human immunodeficiency virus (HIV) is a lentivirus member of the retrovirus family) that causes acquired immunodeficiency syndrome (AIDS), a condition in humans in which the immune system begins to fail, leading to lifethreatening opportunistic infections. Infection with HIV occurs by the transfer of blood, semen, vaginal fluid, pre-ejaculate, or breast milk. Within these bodily fluids, HIV is present as both free virus particles and virus within infected immune cells. The four major routes of transmission are unsafe sex, contaminated needles, breast milk, and transmission from an infected mother to her baby at birth (Vertical transmission) Screening of blood products for HIV has largely eliminated transmission through blood transfusions or infected blood products in the developed world.HIV primarily infects vital cells in the human immune system such as helper T cells (specifically CD4+ T cells), macrophages, and dendritic cells. HIV infection leads to low levels of CD4+ T cells through three main mechanisms: firstly, direct viral killing of infected cells; secondly, increased rates of apoptosis in infected cells; and thirdly, killing of infected CD4+ T cells by CD8 cytotoxic lymphocytes that recognize infected cells. When CD4+ T cell numbers decline below a critical level, cell-mediated immunity is lost, and the body becomes progressively more susceptible to opportunistic infections. Epidemiology UNAIDS and the WHO estimates that AIDS has killed more than 25 million people since it was first recognized in 1981, making it one of the most destructive pandemics in recorded history. Despite recent improved access to antiretroviral treatment and care in many regions of the world, the AIDS pandemic claimed an estimated 2.8 million (between 2.4 and 3.3 million) lives in 2005 of which more than half a million (570,000) were children. In 2007, between 30.6 and 36.1 million people were believed to live with HIV, and it killed an estimated 2.1 million people that year, including 330,000 children; there were 2.5 million new infections. Current scenario in India India’s first known HIV infection was diagnosed in a female sex worker in Chennai in February 1986. It is highly probable that HIV had been circulating for some years before that, since screening during 1986-87 found as many as 3%4% of sex workers infected in Vellore and Madurai, and 1% of STD patients infected in Mumbai. As there were already over 20,000 cases in the world before any case was identified in India, screening for HIV infections began in India in 1985, almost as soon as tests for the HIV antibody were available. Across the world, country estimates have been revised with better data through community based and population surveys. In India, too, for the first time in 2006, HIV testing was a part of the National Family Health Survey (NFHS). The results of NFHS-III give us more accurate information about the estimates of those infected with HIV in the country. Materials and method Selection of patients and collection of data Data and blood collection was done from various hospitals and pathology laboratories of Gujarat . Samples form a total of 58 patients, including 16 Biostatistical analysis of the HIV-aids patients with reference to various factors International Journal of Parasitology Research, ISSN: 0975-3702, Volume 1, Issue 2, 2009 16 women, aged between 15-50 years (median = 33), These included patients suffering from HIV AIDS was collected. Patients identified as suffering from HIV were classified according to the results of the ELISA and western blot detection with the doctors’ guidance. A data of their age, gender, BMI, family history, hemoglobin and history, mode of infection was generated. ELISA Test Patient’s blood samples was collected and centrifuge the sample at 2000 rpm for 5 min. collect the supernatant comprising mainly of serum. Patients not in window period are those that contains HIV antibody in serum sample which was detected by ELISASPOT test kit from genei (Banglore). Microscopic Examination Differential staining was performed of the samples from all 58 patients. For this the morning sputum of patients was collected and were examined by acid fast staining for the detection of mycobacterium tuberculosis infection. Procedure for acid-fast staining Prepare smear from sputum specimen on glass slide, hit fix the smear. Flood the smear with carbol – fuchin stain. Heat gently and avoid boiling when steam arises. wash the slide with water. Cover slide with 20% sulfuric acid for 1 min, then counter stain with methylene blue for 1 min. Examine microscopically under oil immersion lenses. For further conformation fluorescent stain and florescent microscope was used. Analysis of data: Biostatistical analysis was done of the data. Bar chart analysis was carried out of the collected data. Results Data analysis of samples with respect to different factors affecting HIV was done as shown in table 1. The collected samples of HIV affected patients showed a male is to female ratio of 3:1, where 70% of the patients were aged between of 15-40. This age showed maximum prevalence of HIV. The study of the BMI ratio showed that 65% of the patients were thin or normal with no patients showing high BMI ratio (corresponding to thin body built). HIV 1 0 10 20 30 40 50 60 70

Oncoproteomics: New trends in analytical techniques

Oncoproteomics is the study of proteins and their interactions in a cancer cell by proteomic technologies and has the potential to revolutionize clinical practice, including cancer diagnosis. Oncoproteomics screening based on proteomic platforms as a complement to histopathology, individualized selection of therapeutic combinations that target the entire cancer-specific protein network, real-time assessment of therapeutic efficacy and toxicity and rational modulation of therapy based on changes in the cancer protein network associated with prognosis and drug resistance. Oncoproteomics refers to the application of proteomic technologies in oncology and parallels the related field of oncogenomics. The challenges ahead and perspectives of oncoproteomics for biomarkers development are also addressed. With a wealth of information that can be applied to a broad spectrum of biomarker research projects serves as a reference for biomarker researchers, scientists working in proteomics and bioinformatics, oncologists, pharmaceutical scientists, biochemists, biologists, and chemists. Keywords - Oncoproteomics, Genomics, Proteomics, NMR, MS, GC

Immunotherapy and Therapeutic antibodies

Treatment of disorders through immune manipulation of the host has come a long way in the last decades and has broadened its applications from infectious diseases to control of allograft rejection, induction of tolerance for the treatment of autoimmunity and break of tolerance to induce cancer rejection. Immunotherapy, biological response modifier therapy or biotherapy uses the immune system to fight disease like cancer. The potentials of immunotherapy are many. The objective of this review article is to increase awareness of contemporary immunologic therapies and recent advances in immunotherapy. Keywords - Immunotherapy, Adjuvant, Interleukin, Monoclonal antibody, Interferon, Autoimmunity Immunotherapy Introduction Immunotherapy uses the immune system to fight disorders and immune based therapies are to harness the sensitivity, specificity and self- regulation of the immune system to eradicate any and all tumor cells (1). Biological response modifiers (BRMs) change the route the bodys defenses interact with infected cells and produced in a laboratory and given to patients to enhance immune system. BRMs include nonspecific immunomodulating agents, interferons, interleukins, colony-stimulating factors, monoclonal antibodies, cytokine therapy, and vaccines. Monoclonal antibodies are agents, produced in the laboratories that bind to infected cells. Examples of monoclonal antibody therapy include trastuzumab for breast cancer and rituximab for lymphoma. Chronic disorders including malignant, infectious and autoimmune conditions are difficult to treat and often require the use of multiple agents, alone or in combination. The latter approach is already being attempted in some cancer patients. Herceptin, which targets cell proliferation mediated by HER2 and bevacizumab, which targets tumor angiogenesis mediated by VEGF, are being used in combination in breast cancer patients (2). Research has been undertaken to provide an explanation on how a relatively limited number of antibodies can protect against a vast and almost infinite number of invading antigens (3). Proposed potential explanations include the notion that one antibody may be capable of binding more than one antigen through the selection of specific binding sites on pre-existing antibody conformations, and in this manner antibodies with multiple specificities can be achieved. It has been suggested that multispecificity may evolve and play a role in the efficient antibody repertoire for immune protection with one antibody performing more than one task (4). Autoimmune beneficial from the use of two-in-one antibodies and autoimmune disorders, particularly rheumatoid arthritis and systemic lupus erythematosus, are being treated with single biologic agents directed at specific pathogenic pathways, but one may envision the use of two-in-one antibodies directed at multiple targets in the not too distant future (5). Cytokine therapy is used with advanced melanoma and with adjuvant therapy and reaches all parts of the body to kill infected cells and prevent tumors from growing. Vaccine therapy involves use of vaccines produced in Escherichia coli , yeast or insect cells. Plant- based vaccines are also advantageous in terms of their scalability, no cold chain required, stability, safety, cost-effectiveness and needle- free administration (6). Peptide immunotherapy using dominant T-cell epitopes derived from allergens represents another approach to developing safer treatment, because regions involved in allergenicity i.e., responsible for reactivity with IgE, can be completely removed while retaining immunogenicity (7). Antigen- specific immunotherapy strategies include administration of: proteins/peptides with or without adjuvant; dendritic cells (DCs; transfected with DNA- or RNA-encoding tumor antigens, or loaded with peptides, whole proteins or tumor lysates); recombinant viruses encoding tumor antigens; and autologous or allogeneic tumor cells (8). Future of immune therapy for cancer holds promise with novel combined approaches that simultaneously target cancer-initiating stem cells, restore APC immune-stimulating activity, expand tumor-reactive T cells and down regulate suppressor pathways to generate effective therapy.(9) Immune therapy for lung cancer has high potential. Identification of the cell type capable of sustaining neoplastic growth and directing immune therapy to cells that possess