Home
Resources
Publications
Large-scale Production, Bacterial Localization Assessment and Immobilized Metal Affinity Chromatography Purification of a Human Single-Chain Fv Antibody Against alphaIIb-beta3 Integrin

Large-scale Production, Bacterial Localization Assessment and Immobilized Metal Affinity Chromatography Purification of a Human Single-Chain Fv Antibody Against alphaIIb-beta3 Integrin

R Robert, G Clofent-Sanchez, A Hocquellet, M-J Jacobin-Valat, D Daret, A M Noubhani, X Santarelli

Int J Biol Macromol. 2006 Aug 15;39(1-3):51-9.

PMID: 16620955

Abstract:

Our objective was to investigate the Escherichia coli localization (such as supernatant, cytoplasm and inclusion bodies) of an anti-alphaIIb-beta3 (alphaIIbbeta3) scFv fragment referred to as scFv[EBB3] produced in batch fermentation. Immobilized metal affinity chromatography (IMAC) purification was performed on supernatant using expanded bed absorbed technology (EBA) and on sonicated cells in native conditions over an immobilized copper-ion affinity column. Inclusion bodies were solubilized before IMAC purification and the refolding procedure was performed on the column. The majority of scFv[EBB3] were present as inclusion bodies (55%), whereas 36% were found in the cytoplasm and only 9% secreted in the supernatant. The scFv activity was assessed by enzyme-linked immunosorbent assay (ELISA), flow cytometry and immunohistochemistry analyses performed on a thrombus induced in vivo on an atherosclerotic rabbit model.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42417460	Anti-phospho-Integrin β3 (pTyr773) antibody produced in rabbit			Price

As a specialist in analytical chemical Products, Alfa Chemistry offers consumers a wealth of raw materials and a full range of technologies and services.

QUICK LINKS

HEADQUARTERS

Tel:

Fax:

Email: