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- Title
Efficient Conjugation of Rabbit Fab' with β-D-Galactosidase from <em>Escherichia coli</em>.
- Authors
Yoshitake, S.; Hamaguhi, V.; Ishikawa, E.
- Abstract
An efficient procedure for the conjugation of rabbit Fab' with β-D-galactosidase from Escherichia coli using N',N'-w-phenylenedimaleimide is described. Thiol groups of Fab' were stabilized by the presence of ethylenediaminetetraacetate, and maleimide groups were shown to be stable at pH 5 at 4°C. The stability of thiol and maleimide groups enabled an efficient introduction of maleimide groups into Fab' and the average number of maleimide groups introduced into Fab' was 0.76 (range 0.73-0.79; n 10) per molecule. As a result. 43.4% (range 41.3-46,9%; n=6)of Fab' used could be conjugated with most of β-D-galactosidase used. The average number of Fab' molecules conjugated per enzyme molecule was calculated to be 4.2 (range 4.0-4.6; n=6). Both the enzyme and antibody activities were well preserved in the conjugate. There was no self-coupling of Fab', although the enzyme was polymerized to some extent during the conjugation reaction. The enzyme activity and cross-link in the conjugate was stable at pH 6.0-7.0 at 4°C for at least 3 months.
- Subjects
HYDROGEN-ion concentration; ACIDITY function; BUFFER solutions; ENZYMES; ESCHERICHIA coli; ENTEROBACTERIACEAE; MOLECULES; AVOGADRO'S law; BIOMOLECULES
- Publication
Scandinavian Journal of Immunology, 1979, Vol 10, Issue 1, p81
- ISSN
0300-9475
- Publication type
Article
- DOI
10.1111/j.1365-3083.1979.tb01338.x