[Development of monoclonal antibody-based immunoenzyme kit for detection of human thyrotropin hormone]. / Razrabotka immunofermentnogo nabora dlia opredeleniia tireotropnogo gormona cheloveka na osnove monoklonal'nykh antitel.

An enzyme immunoassay kit based on monoclonal antibodies (Mab) has been developed for measuring thyroid-stimulating hormone in human serum and plasma. Laboratory trials demonstrated its high sensitivity (3.1 microIU/ml), specificity, reliability, and accuracy. The analysis is simple and rapid (2.5 h), which is convenient for clinical use. The kit permits differentiation between normal subjects and patients with primary hypo- or hyperthyroidism and helps monitor the treatment efficacy. Comparison with the Amerlite TSH-60 kit, carried out on 357 sera, showed a high correlation coefficient (0.96) between the two kits.

[Immunochemical and functional study of the latrotoxin molecule]. / Immunokhimicheskoe i funktsional'noe issledovanie molekuly latrotoksina.

A panel of monoclonal antibodies (mAb) against alpha-latrotoxin (LT) has been produced and their main characteristics have been determined. The influence of mAb on the functional effects of LT in synaptosomes from rat brain and on the channel formation in bilayer lipid membrane has been investigated. These mAbs do not inhibit binding of LT to rat synaptosomes but modify LT-receptor interaction in terms of LT's channel-forming and secretogenic effects. Antibodies A6 and A24 block these effects, whereas A4 partially preserves the secretogenic action of LT and completely blocks its channel-forming action. Only antibodies A15 affect the LT ability to form cationic channels in BLM, inducing considerable decrease in the frequency of the channel formation. These data and their analysis allow to identify several functional (and, probably, structural) domains of LT responsible for: 1) toxin-receptor interaction; 2) channel-forming and related calcium-dependent secretogenic effects; 3) calcium-independent secretogenic effects; 4) formation of cationic channels in the artificial lipid bilayer.

[Identification and isolation of the protein insect toxin (alpha-latroinsectotoxin from venom of the spider Latrodectus mactans tredecimguttatus]. / Identifikatsiia i vydelenie belkovogo insektotoksina (alpha- latroinsektotoksina) iz iada pauka Latrodectus mactans tredecimguttatus.

The crude venom of spider Latrodectus mactans tredecimguttatus was fractionated by the combination of anion exchange and hydrophobic chromatography. The biological activity of fraction was tested by means of: 1) estimation of toxicity for housefly larva; 2) intracellular recording of miniature excitatory potentials (MEPSPs) in blowfly larvae muscle fibres. As a result of sequential procedures of chromatography separation a homogeneous protein of 120 kilodalton molecular weight was obtained. This protein referred to alpha-latroinsectotoxin produced: 1) a great increase of the frequency of MEPSPs in the dose of 4.2.10(-10) M and its paralytic dose for fly larva was approximately 20 ng/species; 2) no influence of the MEPSPs after application in the dose of 1.2.10(-7) M to the neuromuscular junction of the frog.

[Isolation of alpha-latrotoxin from karakurt spider venom using monoclonal antibodies]. / Vydelenie alpha-latrotoksina iz iada pauka karakurta s pomoshchiu monoklonalnykh antitel.

A method of alpha-latrotoxin (LT) isolation from the venom of Latrodectus mactans tredecimguttatus by means of immunoaffinity chromatography on sepharose conjugated with monoclonal antibodies against LT has been developed. This one-step, high-yield, relatively simple and rapid procedure yields active LT for structural and functional studies of its receptor.