Andrew Ingram

SERRS immunoassay for quantitative human CRP analysis.

SERRS has been used for the first time for the measurement of C-reactive protein (CRP) in an immunoassay. CRP, a biological marker for the diagnosis of infection and inflammation, is quantified in an ELISA using conventional reagents, but the usual colorimetric detection step is replaced by SERRS detection, offering improved sensitivity and potential for multiplexing analysis.

Quantitation of biomolecules conjugated to nanoparticles by enzyme hydrolysis

A novel method for quantifying biomolecules immobilised onto gold and silver nanoparticles is reported; fluorescent-labelled antibodies and DNA are hydrolysed on the surface of the nanoparticles by the addition of trypsin and DNase I, respectively, resulting in the release of the quantifiable fluorescent label into the bulk solution.

Isolation of recombinant proteins from culture broth by co-precipitation with an amino acid carrier to form stable dry powders.

Protein‐coated microcrystals can be generated by co‐precipitation of protein and a water‐soluble crystalline carrier by addition to excess water miscible organic solvent. We have investigated this novel process for its utility in the concentration and partial purification of a recombinant protein exported into the culture broth during expression by Pichia pastoris. Co‐precipitation with a L‐glutamine carrier selectively isolated the protein content of the culture broth, with a minimal number of steps, and simultaneously removed contaminants including a novel yeast metabolite. This pigment co‐elutes during aqueous chromatography but its elucidation as a benzoylated glycosamine suggested a simple route of removal by partition during the co‐precipitation process. Scale‐up of the process was readily achieved through in‐line mixing and subsequent reconstitution of the dried protein‐coated microcrystals yielded natively folded, bioactive protein. Additional washing of the crystals with saturated L‐glutamine facilitated further purification of the recombinant protein immobilized on the L‐glutamine carrier. Thus, we present a novel method for the harvesting of recombinant protein from culture broth as a dry powder, which may be of general applicability to bioprocessing. Biotechnol. Bioeng. 2010;106: 764–773.

Functionalised nanoparticles and SERRS for bioanalysis

Functionalised nanoparticles have been used in a number of studies including detection of DNA at ultra low concentrations, immuno-histochemistry and more recently as substrates for surface enhanced resonance Raman scattering (SERRS) based imaging approaches. The advantages of using metallic nanoparticles are that they are very bright in terms of their optical characteristics and also can be functionalised to provide a SERRS response and hence provide a unique Raman fingerprint. Here we present the functionalisation of gold and silver nanoparticles in such a way that the enhancement effect can be greatly increased through biological interaction and as such effectively turn on the SERRS effect. In an advancement of this nanoparticles have been used as imaging agents for single cells when functionalised with an appropriate antibody and can give information on the expression of specific receptors on cell surfaces as well as sub-cellular compartmentalisation information.