Production of 'neometalloenzymes' by de novo biosynthesis: New ELISA method for their characterization

E. Nedonchelle, C. Leduc, X. Garcia, Roberto Z Guzman, M. A. Vijayalakshmi

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Several approaches known for producing 'neometalloenzymes' are classified into two categories: protein engineering using antibodies as starting materials and 'de novo' biosynthesis of metal-binding antibodies with potential catalytic metal-binding structure. This latter approach is chosen in this study. Polyclonal anti-zinc-iminodiacetate [IDA-Zn(II)] antibodies are produced in rabbits and mice. Because of the absolute need for the unequivocal screening of the hapten [IDA-Zn(II)] specific antibodies, a new ELISA method was developed using a biheaded polyethylene glycol with biotin on one end and the hapten on the other end. The parameters for optimizing the immunization and the ELISA technique are discussed and the method is validated with rabbit and mice sera.

Original languageEnglish (US)
Pages (from-to)106-117
Number of pages12
JournalAnnals of the New York Academy of Sciences
Volume864
DOIs
StatePublished - 1998

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Biosynthesis
Enzyme-Linked Immunosorbent Assay
Antibodies
Haptens
Metals
Rabbits
Immunization
Protein Engineering
Biotin
Zinc
Screening
Mouse
Rabbit
Serum
Proteins
Protein
Polyethylene Glycol

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Production of 'neometalloenzymes' by de novo biosynthesis : New ELISA method for their characterization. / Nedonchelle, E.; Leduc, C.; Garcia, X.; Guzman, Roberto Z; Vijayalakshmi, M. A.

In: Annals of the New York Academy of Sciences, Vol. 864, 1998, p. 106-117.

Research output: Contribution to journalArticle

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