Determination of the Binding Constant Between Bovine Serum Albumin and Its Monoclonal Antibody by Affinity Capillary Electrophoresis with Laser Induced Fluorescence Detection

Abstract

Abstract: The binding constant(K_b) between fluorescein isothiocyanate labeled bovine serum albumin(FITC-BSA) and its monoclonal antibody was determined using the ligand separation mode of affinity capillary electrophoresis(ACE) with laser induced fluorescence detection(LIF). Free FITC-BSA and the complexes were separated by capillary electrophoresis in 7min after incubation. The running buffer was 100 mmol/L HEPE Sat pH7.4, and separation voltage was 30kV. The K_bwas determined to be 4.7×10⁸L/mol using Scatchard plotting method. The influences of incubation time and electrophoresis conditions on determination were also studied.

Key words: Affinity capillary electrophoresis, Laser induced fluorescence detection, Protein, Binding constant

CLC Number:

O657.8

TrendMD:

WANG Qing-Gang, LUO Guo-An, William S. B. Yeung . Determination of the Binding Constant Between Bovine Serum Albumin and Its Monoclonal Antibody by Affinity Capillary Electrophoresis with Laser Induced Fluorescence Detection[J]. Chem. J. Chinese Universities, 1999, 20(10): 1551.

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Determination of the Binding Constant Between Bovine Serum Albumin and Its Monoclonal Antibody by Affinity Capillary Electrophoresis with Laser Induced Fluorescence Detection

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