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Production of monoclonal antibodies specific to human von willebrand factor

Rosely V. Meissner; Israel Roisenberg; Nance B. Nardi

Departamento de Genética, Universidade Federal do Rio Grande do Sul, Caixa Postal 15053, 91501-970 Porto Alegre, RS, Brasil. Send correspondence to I.R.

ABSTRACT

A monoclonal antibody specific for human von Willebrand factor, vW1E2, was produced after immunization of BALB/c mice with partially purified FVIII/von Willebrand factor. Testing of sera and screening of hybridomas was done by ELISA using rabbit anti-vWF serum as capture layer of vWF. The specificity of vW1E2 to vWFAg was determined by testing it against plasmas of normal individuals, hemophiliacs A and vWD patients. This specificity was corroborated by imunoprecipitation experimems in which vWFAg was precipitated from plasma by vW 1E2 with the aid of rabbit anti- mouse Ig serum,' as assessed by electroimmunoassay (EIA) and ELISA. An ELISA employing vW1E2 was evaluated as an assay for quantification of plasmatic vWFAg. Plasma samples from type I and type II vWD patients were analyzed as well as from hemophilic A patients and normal individuals. The levels of vWFAg obtained by ELISA were compared to those by EIA and a significant correlation was observed (r= 0.735; df = 35; P < 0.001).

Keywords: Monoclonal antibodies; Human; Von willebrand factor.

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