Production in Escherichia coli of a functional murine and murine::human chimeric F(ab′) 2 fragment and mature antibody directed against human placental alkaline phosphatase

• Published in: Journal of biotechnology 1995-09, Vol.42 (2), p.133-143
• Main Authors: Flamez, Daisy, Remaut, Erik, Fiers, Walter
• Format: Article
• Language: English
• Subjects: Alkaline Phosphatase - immunology; Animals; Base Sequence; Biomarkers, Tumor - immunology; Biotechnology; Chimeric antibody; Cloning, Molecular; DNA, Complementary; Escherichia coli - immunology; Escherichia coli - metabolism; F(ab′) 2 fragment; Genetic Vectors - chemistry; Genetic Vectors - genetics; Genetic Vectors - metabolism; Humans; Immunoglobulin Fab Fragments - biosynthesis; Immunoglobulin Fab Fragments - genetics; Immunoglobulin G - genetics; Immunoglobulin G - metabolism; Mice; Molecular Sequence Data; Placenta - enzymology; Placental alkaline phosphatase; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/0168-1656(95)00072-X

We report the production in Escherichia coli of a murine antibody IgG2b, a murine::human chimeric antibody IgG3 and the corresponding F(ab′) 2 fragments, all directed against human placental alkaline phosphatase, a tumor marker. The cDNA of the heavy chain of the mature antibodies and their fragments were linked up to the bacterial alkaline phosphatase signal sequence and were placed under control of the inducible tac promoter. Coexpression with the murine κ light chain resulted in production of functional dimeric, trimeric and tetrameric, mature antibodies and F(ab′) 2 fragments in the periplasm of E. coli in a yield of 200–300 μg 1 −1. High amounts of light and heavy chains were present also in the insoluble fraction.