A genetic system to elicit and monitor anti-peptide antibodies without peptide synthesis

Journal Article · Fri Feb 01 00:00:00 EST 1991 · Bio/Technology; (United States)

DOI:https://doi.org/10.1038/nbt0291-170· OSTI ID:5148323

Martineau, P; Charbit, A; Werts, C; O'Callaghan, D; Hofnung, M ^[1]; Leclerc, C ^[2]

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1. Centre National de la Recherche Scientifique UA271, Paris (France)
2. Inst. Pasteur, Paris (France)

The authors present a simple and flexible procedure to elicit and assay anti-peptide antibodies without peptide synthesis. It consists of expressing the peptide of interest in the form of a genetic insert within two different recipient' bacterial proteins. One hybrid protein is used as immunogen for the induction of antibodies against the inserted peptide and the other as antigen for monitoring the anti-peptide antibodies raised. The two recipient' proteins used are the MalE and the LamB proteins from E. coli. The MalE hybrid proteins can be affinity purified on an amylose column using mild nondenaturing conditions and can be crystallized for structural studies; LamB hybrid proteins express the inserted peptide on the cell surface so that intact bacteria can be used as a reagent. The authors chose, as a model peptide, a B-cell epitope from the pre-S(2) region of Hepatitis B virus. With both MalE and LamB hybrid proteins, high titers of anti-preS antibodies, able to react with native HBsAg particles, were induced in mice. The anti-peptide antibody titers recorded by ELISA were comparable to those obtained when either a synthetic peptide, or the hybrid proteins, were used as immobilized antigen.

• OSTI ID: 5148323
• Journal Information: Bio/Technology; (United States), Vol. 9:2; ISSN 0733-222X
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ANTIBODIES
INDUCTION
IMMUNITY
DETECTION
ANTIGENS
BACTERIA
CHROMATOGRAPHY
ESCHERICHIA COLI
GENETICS
HEPATITIS
HYBRIDIZATION
IMMUNOLOGY
PEPTIDES
PROTEINS
VIRUSES
BIOLOGY
DIGESTIVE SYSTEM DISEASES
DISEASES
MICROORGANISMS
ORGANIC COMPOUNDS
PARASITES
SEPARATION PROCESSES
550200* - Biochemistry