Forensic Applications of ELISA

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1. Monoclonal antibodies (MoAbs) to ABO and MN antigens, glycophorin A and B, IgG allotypes G3M G and T, A2-seminoglycoprotein (A2SGP), and paraquat were produced for use in ELISAs.2. Human blood could be identified by inhibition-ELISAs using ALB, IgG, A2M, HP, TF, GC, A2HS, BF, and HF as indicator proteins. Human urinary stains were best identified by a capture ELISA using anti-uromucoid as the capture antibody.3. Automation and data processing for ABO grouping of body fluids were effected by assembly of a reaction unit containing a plate coated with stain-derived, detergent-solubilized antigen, and an automatic reading unit.4. A simple dot-immunobinding (dot-ELISA) method for GM typing was developed using MoAbs to G3M G and G3M T. The method is well suited for routine forensic casework.5. A competitive ELISA for paraquat was developed which enabled determination of 1-10 ng/ml of the pesticide.6. Saliva, semen, and blood in mixed stains could be differentially ABO grouped by capture ELISAs using corresponding antibodies. In particular, home-made MoAbs to AB-carrying A2SGR specifically captured the corresponding protein, thereby enabling correct ABO grouping of semen mixed with vaginal fluids. Thus the method appears useful in rape casework.7. Production of MoAbs to IgG allotypes GlM F and GlM X is in progress, since four GM monoclonals are required for routine GM typing of Japanese.