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Detection of Antiviral Antibodies with Predetermined Specificity Using Synthetic Peptide--Lactamase Conjugates: Application to Antibodies Specific for the preS Region of the Hepatitis B Virus Envelope Proteins

¹ The Lindsley F. Kimball Research Institute of The New York Blood Center, 310 East 67th Street, New York 10021, U.S.A.
and² Division of Biology, California Institute of Technology, Pasadena, California 91125, U.S.A.

Amino acid sequences coded for by the preS region of the hepatitis B virus (HBV) envelope gene are present both in HBV and in subviral hepatitis B surface antigen (HBsAg) particles. Consequently, anti-preS-specific antibodies are elicited during the course of HBV infection. Such antibodies are virus-neutralizing. Therefore, it is important to determine whether or not vaccination with HBsAg also induces an anti-preS-specific immune response. We describe here an enzyme-linked immunosorbent assay applicable for the screening of sera from vaccinated individuals for anti-preS antibodies. IgG from serum specimens was adsorbed to staphylococcal Protein A on a superparamagnetic support and subsequently mixed with a synthetic peptide analogue [preS(120–145)] covalently linked to -lactamase. The presence of anti-preS in serum specimens resulted in binding of the conjugated -lactamase to the magnetic support. The adsorbed enzyme was quantified colorimetrically.

Keywords: HBV, ELISA, preS antibodies, synthetic peptide

The following contributors, listed alphabetically, provided serum specimens from persons vaccinated with different hepatitis B vaccines: L. Baker,¹ J. Desmyter,³ M. Girard,⁴ P. N. Lelie,⁵ H. W. Reesink,⁵ C. E. Stevens,¹ P. Taylor¹ and F. Tron⁴ (³Rega Institute, University of Leuven, Leuven, Belgium; ⁴Pasteur Vaccins, Marnes-La Coquette, France; ⁵Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam, The Netherlands).

Received 13 August 1985; accepted 29 October 1985.

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