University of North Florida
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Stuart Chalk, Ph.D.
Department of Chemistry
University of North Florida
Phone: 1-904-620-1938
Fax: 1-904-620-3535
Email: schalk@unf.edu
Website: @unf

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Alexander P. Osipov

Abbrev:
Osipov, A.P.
Other Names:
Address:
Laboratory of Enzyme Engineering, Chemistry Faculty, M.V. Lomonosov Moscow State University, Russia.
Phone:
[7](095) 939-34-07
Fax:
[7](095) 939-27-42
Email:

Citations 4

"New Immunoassay Technique Using Antibody Immobilized On A Membrane And A Flow Cuvette As Reaction Vessel"
J. Immunol. Methods 1993 Volume 157, Issue 1-2 Pages 11-17
B. M. Gorovits, A. P. Osipov* and A. M. Egorov

Abstract: Immunoenzymatic detection systems have been developed using human IgG as a model antigen. A membrane with covalently immobilized specific antibodies was placed into a specially constructed ultranarrow flow cuvette and solutions containing the antigen and antibody-peroxidase conjugate were then successively passed through the flow capillary cell. After washing, the membrane was placed into the substrate solution and the intensity of developed color on the membrane was recorded visually or by a reflection spectrophotometer. The lower detection limit was about 5 x 10^-11 M and the overall analysis time was 10 min. Photoimmobilization was used to immobilize the antibody and thereby permitting control of the protein surface concentration on the membrane as well as the dimensions and shape of the activated region.
Immunoglobulin G Immunoassay Spectrophotometry Immobilized antibody Photochemistry Membrane

"Studies On Antigen-antibody Interactions By Flow Injection Chemiluminescent Sensor"
J. Biolumin. Chemilumin. 1997 Volume 12, Issue 1 Pages 37-39
Alexander P. Osipov, Natalya V. Zaitseva, Alexey M. Egorov

Abstract: This system was successfully used for analyzing the kinetics of bimolecular interaction peroxidase antibodies against peroxidase and for concentration determination (5). In this paper we report our further study on the determination of kinetic rate constants for processes of bimolecular interactions on the boundary liquid-solid phase (sensor surface) between small haptens (simazine and 2,4-dichlorophenoxyacetic acid (2,4-D)) and specific antibodies using this technique.
Antigens Chemiluminescence Sensor Complexation Kinetic Enzyme

"Flow Injection Immunoassay Of Human IgG In A Kinetic Regime"
Biotekhnologiya 1989 Volume 5, Issue 7 Pages 808-810
Osipov, A.P.;Arefyev, A.A.;Egorov, A.M.

Abstract: A solid-phase enzyme immunoassay of human IgG in the flow-injection mode is described. Samples of 20-50 mL of IgG solutions were injected into a polyethylene tubing leading 0.05 M phosphate buffer (pH 7.4, 0.14 M NaCl) at a flow rate of 0.5 mL/min to a 2 x 50 mm column packed with Sepharose 4B with BrCN-immobilized human IgG and immunologically bound rabbit antibody-horseradish peroxidase complexes. Peroxidase released into the eluate was mixed with H2O2 and ABTS substrates and the absorbance of the resulting colored product was measured in a flow cuvette at 420 nm. The contact between the samples and column packing (stopped flow) was varied 0.5-10 min; the contact between 0.5-2 min yielded almost linear calibration responses. The detection limits were 0.32 and 0.16 mg/mL for the contract time 0.5 and 10 min, respectively. The column allows ~100 samples to be analyzed without peroxidase complex reloading. Successive analyzes of the same sample show ~1% decrease in resulting values per run. The analyzes last 1-2 min each and have a coefficient of variation 3.2%. (SFS)
Immunoglobulin G Immunoassay Kinetic Stopped-flow Immobilized enzyme Sepharose beads

"Chemiluminescent Immunoenzyme Biosensor With A Thin-layer Flow-through Cell. Application For Study Of A Real-time Bimolecular Antigen-antibody Interaction"
Biosens. Bioelectron. 1996 Volume 11, Issue 9 Pages 881-887
Alexander P. Osipov*, Natalia V. Zaitseva and Alexey M. Egorov

Abstract: A simple flow enzyme system for real-time continuous monitoring of interaction of biological molecules has been developed. It relies upon a thin-layer flow-through cell placed directly into the measuring compartment of the luminometer. One ligand (antibody) is immobilized on the inner surfaces of the flow cuvette, and a second ligand (antigen) labeled with a peroxidase molecule moves through the flow cell. The quantity of the complex on the surface of the cell may be monitored by measurement intensity of chemiluminescence in the reaction of peroxidase label with substrates (p-iodophenol, luminol and hydrogen peroxide). In such a way one can detect in a real-time regime the kinetics of association (or dissociation) of the complex labeled ligand- receptor on the surface of the cuvette. Due to the small thickness of the flow cell the diffusion limitations of interaction for two kinds of biomolecules (soluble and immobilized) are negligible, so the resulting intensity of chemiluminescent signal reflects the kinetics of interaction between soluble and immobilized components. The system may be successfully used for molecular recognition studies, analyzing the kinetics of bimolecular interaction and for concentration determination.
Chemiluminescence Sensor