Immobilized metal-ion affinity chromatography of human antibodies and their proteolytic fragments

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Daniela Todorova-BalvayM Vijayalakshmi

Abstract

Immobilized metal-ion affinity chromatography (IMAC) performed with four different transition metal ions: copper(II), nickel(II), zinc(II) and cobalt(II), was used to study the adsorption properties of human polyclonal gamma-globulines (IgG), Cohn II-III fractions, and their pepsin cleaved fragments: Fab'2 and F'c. In each case, digested products showed lower affinity for metal ions, as well by decreasing pH elution as by competition with imidazole. An explanation was proposed by the presence of a histidine (His) cluster in the F'c domain of IgGs, identified by computer calculation (accessible surface area (ASA) determination) as the more probable His 433-x-His 435 sequence presented in the CH3 domain of human IgG heavy chain. As shown by IMAC and electrophoresis, F'c and undigested IgG have higher affinity for transition metal ions than Fab'2 fragments and could be then separated in one step by IMAC. When chelated Zn(II) or Co(II) are used as ligands, the Fab'2 fragment could be easily recovered under mild conditions (pH 7) in the non-retained fraction. This approach could be used as a powerful alternative to conventional protein A/G methods for the commercial preparation of non immunogen active Fab'2 fragments.

Citations

Mar 26, 2011·Inorganic Chemistry·Daniele SannaEugenio Garribba
Oct 29, 2011·Methods : a Companion to Methods in Enzymology·B Vijayalakshmi AyyarRichard O'Kennedy
Nov 11, 2011·Journal of Chromatography. a·Pete Gagnon
Apr 27, 2010·Journal of Chromatography. a·Rajasekar R Prasanna, Mookambeswaran A Vijayalakshmi
Mar 15, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Cecília Alves MourãoSonia Maria Alves Bueno
Feb 12, 2008·Journal of Biochemical and Biophysical Methods·Pengli BaiChangsheng Zhao
Dec 11, 2007·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Mariana Borsoi RibeiroSonia Maria Alves Bueno
Jul 10, 2007·Journal of Chromatography. a·Ana C A RoqueM Angela Taipa
Feb 20, 2007·Analytical Biochemistry·Ben W MuirN Joe Maeji
Jul 16, 2015·Biotechnology and Bioengineering·Alexander KinnaKerry Chester
Dec 13, 2006·Journal of Separation Science·Petr PrikrylZdenka Kucerová
Jul 13, 2006·Journal of Separation Science·Eva Zatloukalová, Zdenka Kucerová
Aug 10, 2006·Chembiochem : a European Journal of Chemical Biology·Rafael A VegaChad A Mirkin
May 19, 2006·Biomedical Chromatography : BMC·Hongqin FengXicheng Wang
Sep 3, 2013·Journal of Molecular Recognition : JMR·Mariana Borsoi-RibeiroSônia Maria Alves Bueno
May 24, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Chaoji HuangfuJingang Zhang
Oct 8, 2005·Expert Review of Proteomics·Xuesong SunQing-Yu He
Dec 26, 2016·Methods : a Companion to Methods in Enzymology·Sushrut AroraB Vijayalakshmi Ayyar

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.