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A simple method for midkine purification by affinity chromatography with a heavy chain variable domain (VH) fragment of antibody.

W Dansithong·S Paul·Y Kojima·K Kamiya·T Shinozawa
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Abstract

A DNA fragment for a heavy chain variable domain (VH) was prepared from a hybridoma that produces a monoclonal antibody against human midkine (MK). The antibody fragment was produced in Escherichia coli and its affinity for chemically synthesized full length MK or recombinant midkine c-terminus (MKc-half) protein was confirmed by ELISA. An Escherichia coli cell lysate expressing MKc-half was applied to a VH fragment-coupled Sepharose 4B column and eluted with a buffer containing 0.5 M NaCl. SDS-PAGE analysis revealed a high degree of purity of the MKc-half protein in the eluent, showing the utility of a recombinant VH fragment in purification of proteins by affinity chromatography.

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Amino Acid SequenceAntibodies, MonoclonalBase SequenceCarrier ProteinsChromatography, AffinityCytokinesEscherichia coliGenetic VectorsImmunoglobulin Heavy ChainsImmunoglobulin Variable RegionMidkineMolecular Sequence DataRecombinant ProteinsJournal ArticleResearch Support, Non-U.S. Gov't

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W Dansithong, S Paul, Y Kojima, K Kamiya, T Shinozawa. 2001-03-28. A simple method for midkine purification by affinity chromatography with a heavy chain variable domain (VH) fragment of antibody.. https://doi.org/10.1016/s0165-022x(00)00145-7

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