Purification of recombinant α-amylase with immuno-affinity chromatography using monoclonal antibody

Masamichi Kamihira, Masayuki Taniguchi, Shinji Iijima, Takeshi Kobayashi

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

A monoclonal antibody against recombinant thermostable α-amylase produced by Escherichia coli was isolated from serum-free medium and immobilized on Sepharose 4B. The adsorption equilibrium between α-amylase and the immobilized immuno-adsorbent showed a Langmuir type isotherm. The breakthrough curve calculated numerically using the averaged volumetric coefficient coincided well with the experimental data. More than 90% of the activity of bound α-amylase could be recovered by eluting with glycine-HCl buffer (pH 2.5). The elution profile at pH 2.5 became sharper with increasing temperature. By using an immuno-affinity column, the recombinant α-amylase produced by E. coli could be purified homogeneously from crude extract enzyme solution with two-step elution.

Original languageEnglish
Pages (from-to)625-631
Number of pages7
JournalJournal of Fermentation Technology
Volume66
Issue number6
DOIs
Publication statusPublished - 1988
Externally publishedYes

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