Immunoenzyme Assessment of Human ApoB-Lipoprotein Binding to Immobilized Receptor of Low Density Lipoproteins. 1. Preparation of Anti-Receptor Monoclonal Antibodies

V. P. Tsibulsky,¹ V. V. Yakushkin,^1,2 O. Yu. Tikhomirov,¹ and S. N. Preobrazhensky³

¹Institute of Experimental Cardiology, Russian Cardiology Complex, ul. 3-ya Cherepkovskaya 15a, Moscow, 121552 Russia; fax: (7-095) 414-66-99.

²To whom correspondence should be addressed.

³USANA Research Company, Salt Lake City, Utah, USA.

Submitted February 19, 1997.

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The preparation and properties of V5 monoclonal antibody to low density lipoprotein receptor (LDL-receptor) from bovine adrenal cortex membranes are described. The monospecific V5 antibody recognizes the LDL-receptor (the only protein with molecular mass of 140 kD) in bovine adrenal cortex membranes. V5 antibody fails to compete with human low density lipoproteins (LDL) for binding to the LDL-receptor. After absorption in standard 96-well polystyrene plates, V5 antibody efficiently binds the affinity-purified LDL-receptor from the solution and the subsequent binding of the LDL-receptor with human LDL was determined using peroxidase-labelled antibodies to apolipoprotein B. The LDL-receptor immobilized by V5 antibodies is suggested for use in studies on the binding of human lipoproteins to the LDL-receptor.

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KEY WORDS: lipoproteins, receptor of low density lipoproteins, monoclonal antibodies, immunoenzyme analysis.

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