[Selection of optimal conditions for synthesis of protein A-laccase conjugates and their use in different variants of immunoenzyme analysis]. / Podbor optimal'nykh uslovii sinteza kon''iugatov belok A-lakkaza i ikh ispol'zovanie v razlichnykh variantakh immunofermentnogo analiza.

A study to determine optimal molar ratios of Protein A to laccase for the synthesis of their conjugate by periodate method is presented. No loss of enzymatic or immunological activity of the conjugate developed was observed during 6 month. The conjugate could be effectively used in various techniques of enzyme immunoassay (competitive or sandwich techniques, dot immunoblotting) for immunoglobulin G. The detection limits of the assays for immunoglobulins were better than 1 ng/ml.

[Immunoenzyme analysis based on laccase conjugates with fluorometric detection of the enzymatic reaction product]. / Immunofermentnyi analiz na osnove lakkaznykh kon''iugatov s fluorimetricheskoi detektsiei produkta fermentativnoi reaktsii.

The possibility of using homovanillic acid as a substrate of laccase (produced by the basidiomycete Coryolus hirsutus) has been demonstrated for the first time. The reaction was shown to result in the formation of a fluorescent product. Several kinetic parameters and optimal conditions were determined for this enzymatic reaction. The use of homovanillic acid as the substrate was found to increase the enzyme immunoassay sensitivity by an order of magnitude, compared to conventional substrates.

[The effect of conditions of synthesis of immunolaccase conjugates on characteristics and composition of the compounds obtained]. / Vliianie uslovii sinteza immunolakkaznykh kon''iugatov na kharakteristiki i sostav poluchaemykh soedinenii.

Optimal conditions for preparing laccase conjugates by the periodate method have been selected. The effect of the initial molar ratio of IgG to laccase and pH of the medium on the composition of laccase conjugates was studied by the HPLC method. The maximum yield of the conjugates was observed, when laccase was oxidized with 0.12 M sodium periodate the pH of the medium was 8.5, and the initial molar ratio of IgG to laccase was 2:1. The conjugates can be stored for one year without any loss in immunological activity.