Study on the Genetic Polymorpphisms of Serum Proteins from Protease-treated Samples and its Application to Forensic Medicine.

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We studied the genetic polymorphisms of serum proteins by using electrophoresis, isoelectric focusing and SDS-PAGE.Samples for the typings were pre-treated with protease(carboxypeptidase B, carboxypeptidase Y, papain, V8-protease, etc.) and sialidase. Patterns after electrophoresis were visualized by electroblotting or immunofixation. The results obtained so far are as follows. 1. We found additional new variants of C3 in a Japanese pupulation. One variant was named as FO2 from a comparative study, and an amino acid substitution of Arg to Gln was found in the FO2 molecule. (submitted for publication). 2. C4 typing was performed successfully from plasma or serum samples. The samples were treated with both carboxypeptidase B and sialidase prior to electrophoresis. The C4 polymorphism is invaluable not only for the paternity test in medico-legal field but also for the study of etiology in clinical medicine. We examined the samples from patients with IgA-nephropathy, pemphigus, bullous pemphigoid or systemic lupus erythematosus, and found that the silent allele of Alocus(C4**AQO) was strongly associated with those diseases.(in press). 3. We found one common and four variant alleles of ATIII system in Japanese. Typing method, variants and nomenclature(alpha-numeric code) were described in our report (in press). 4. In the IEF-patterns of complement HF, many non-genetically controlled bands were observed ; the further study is required. Severa new findings have been obtained during the typings of complement BF and Cls (the results will be describled elsewhere). 5. Detection of genetic marker has been performed from bloodstain sample. The treatment of bloodstain extracts with florisil gives good results for HP and TF typings. We are now studying the determination of genetic markers from bloodstains by using the florisil-and enzyme-trated extracts as the samples of electrophoresis.