Benjamin W. Grunbaum

Application of an improved microelectrophoresis technique and immunoelectrophoresis of the serum proteins on cellulose acetate

Abstract Applications of a recently described adaptation of a microelectrophoresis technique and equipment to use with a cellulose acetate membrane in the study of blood protein fractions are described. The properties of the membrane, properly utilized, allow much better results to be obtained than with filter paper and exhibit technical and practical advantages over gels. The application of the material for rapid immunoelectrophoresis is described. For this purpose, it is markedly superior to gel supports in speed and operational facility. When employed with the technique described, it yields approximately the same degree of definition of immunologically distinguishable fractions as that of gel supports.

Distribution of Gene Frequencies and Discrimination Probabilities for 22 Human Blood Genetic Systems in Four Racial Groups

Gene frequencies were computed in four racial categories from 5956 blood donors from California, Hawaii, Mexico City, and Texas. Calculations were based on the phenotypic distribution of 22 blood genetic systems including 7 blood groups and 15 genetically controlled polymorphic proteins and enzymes. Matching probabilities for 20 systems were approximately 1 in 100 000 Asians, 1 in 200 000 blacks, 1 in 330 000 Mexicans, and 1 in 1 000 000 whites. The complementary discrimination probability, which measures the likelihood that two random individuals do not match, was, for practical purposes, unity. The combined new technology for blood grouping and electrophoresis using cellulose acetate membranes provides a powerful individualizing and discriminating tool for forensic science investigation.

Quantitative analysis of normal human serum proteins on permanently transparentized cellulose acetate membranes

Abstract Reference values for the serum protein distribution for a normal population of the Caucasian male were established. The method employed was that of electrophoresis on cellulose acetate in a Beckman microelectrophoresis apparatus. Following electrophoresis, the supporting cellulose acetate membranes were permanently transparentized and the protein fractions quantitated in a modified Beckman Analytrol.

A micro procedure for fast typing of the genetic variants of phosphoglucomutase.

A fast, sensitive, and specific micro procedure for the determination of the genetic variants of phosphoglucomutase from red blood cell hemolysates is described. Cellulose acetate is used as the supporting medium for electrophoresis. A multiple sample applicator places up to eight samples on the membrane in one operation. Following electrophoresis, the cellulose acetate membrane is inverted onto a precast gel containing a reaction mixture which yields an insoluble colored end product on incubation at 37°C. The whole procedure, including electrophoresis and staining, is complete in 6o–8o minutes. The resolution of the various isoenzymes is adequate and permits ready discrimination of the three common genetic types of phosphoglucomutase, namely types 1, 2–1 and 2.

An automatic one- to eight-sample applicator for fast qualitative and quantitative microelectrophoresis of plasma proteins, hemoglobins, isoenzymes, and cross-over electrophoresis

Abstract An electrophoresis sample application system was developed which provides for fast, reliable, and economical microanalyses to measure both quality and quantity of many blood proteins. The system consists of a flexible eight-sample applicator with a well integrated complimentary 32-sample holder and cell cover. The system permits simultaneous pickup and transfer of one to eight samples and the concurrent electrophoresis of up to 24 specimens on a single cellulose acetate membrane or an agarose gel supporting medium. Examples and illustrations of the quality of analytical results for the fractionation of plasma proteins, hemoglobins, lactic acid dehydrogenase isoenzymes, lipoproteins, and cross-over electrophoresis are described. The resolution of constituents is adequate for comparative analyses in the forensic science laboratory and for quantification in most medical clinical analyses.

The Probability of Non-discrimination or Likelihood of Guilt of an Accused: Criminal Identification

Abstract The relative efficacy of genetic blood group systems is explored in terms of the probability of non-discrimination. This probability is used to assess the value of anti-sera and blood group systems to exclude falsely accused individuals. In criminal identification situations, it is often useful to assess the likelihood that a non-excluded defendants blood was found at the scene of a crime. The probability that an accused individual who matches a specific set of blood phenotypes was present at a crime scene is derived and discussed. These calculations are illustrated with examples using blood group data from up to 5000 samples from blood donors.

Individuality of human serum by immunoelectrophoresis.

A method is described which implements the direct comparison of serum samples from different individuals by simultaneous electrophoresis followed by simultaneous diffusion and precipitation by anti-human horse serum. Results have been evaluated, with an arbitrary reference grid, by comparison of the numbers of precipitin bands present in the paired immunoelectrophoretograms Graphic representation of these differences distinguishes serum samples of like from those of unlike origin.

Studies of primate protein variation and evolution: microelectrophoretic detection.

Genetic variation at 16 protein and enzyme loci in Cercopithecus aethiops and several other primate species has been surveyed, using cellulose acetate microelectrophoresis. Resolution of several standard variant proteins is comparable to that achieved on starch gel or polyacrylamide gel. Although both intraspecific and interspecific variation was observed for some loci, the data generally support the concept that extracellular proteins are more likely to be polymorphic within a species, while intracellular proteins generally vary between species, if at all. These methodologies are particularly appropriate for screening multiple-locus variation in large numbers of samples; their relevance to studies of molecular evolution and evaluation of theories of kin selection is discussed.

The Probability of Exclusion or Likelihood of Guilt of an Accused: Paternity

Abstract The relative efficiency of genetic blood group systems to exclude individuals is discussed. The potential for exclusion based on the combination of genetic systems is calculated showing the utility of genetically controlled polymorphic enzyme/protein and immunologic systems. The key to assessing the guilt or innocence in a paternity dispute using genetic evidence is the estimation of the probability of not being excluded by chance alone. This probability is computed under two sets of conditions: a random male supplied the genes in question, or a male with a phenotype identical to the accused supplied the genes in question. Contrasting these two probabilities leads to an estimate of the likelihood of guilt or innocence associated with the evidence from blood group genetic systems. The presentation gives two tables for the calculation of these non-exclusion probabilities in paternity disputes for currently used biochemical and immunological systems. The role of Bayes’ theorem is discussed in conjunction with these non-exclusion probabilities.

Phenotyping of Haptoglobin on Gradient Acrymalide Gel Slabs using the Beckman Microzone System

A phenotyping procedure for haptoglobin is described utilizing gradient acrylamide gels. The haptoglobin phenotypes (seen as complexes with hemoglobin) are separated by wide margins. In addition, each haptoglobin type shows a unique pattern. At this time, the gradient acrylamide gel electrophoresis results in an unmistakable positive identification of the haptoglobin phenotype. A detailed analytical micro procedure is described for the preparation, electrophoresis, and the preservation in the dry form of the acrylamide gel electrophoretograms.