Considerations in producing preferentially reduced half-antibody fragments

Journal of Immunological Methods
Asta MakaraviciuteAlmira Ramanaviciene

Abstract

Half-antibody fragments are a promising reagent for biosensing, drug-delivery and labeling applications, since exposure of the free thiol group in the Fc hinge region allows oriented reaction. Despite the structural variations among the molecules of different IgG subclasses and those obtained from different hosts, only generalized preferential antibody reduction protocols are currently available. Preferential reduction of polyclonal sheep anti-digoxin, rabbit anti-Escherichia coli and anti-myoglobin class IgG antibodies to half-antibody fragments has been investigated. A mild reductant 2-mercaptoethylamine (2-MEA) and a slightly stronger reductant tris(2-carboxyethyl)phosphine (TCEP) were used and the fragments obtained were quantitatively determined by SDS-PAGE analysis. It has been shown that the yields of half-antibody fragments could be increased by lowering the pH of the reduction mixtures. However, antibody susceptibility to the reductants varied. At pH4.5 the highest yield of sheep anti-digoxin IgG half-antibody fragments was obtained with 1M 2-MEA. Conversely, rabbit IgG half-antibody fragments could only be obtained with the stronger reductant TCEP. Preferential reduction of rabbit anti-myoglobin IgG antibodies was opt...Continue Reading

References

Mar 15, 1976·Clinica Chimica Acta; International Journal of Clinical Chemistry·K MiyaiY Kumahara
Aug 1, 1963·Proceedings of the National Academy of Sciences of the United States of America·J L PALMERK E VANHOLDE
Oct 20, 2005·Journal of Bioscience and Bioengineering·K NakanishiK Imamura
Apr 17, 2007·Analytical Biochemistry·Il-Hoon ChoSe-Hwan Paek
Nov 16, 2011·The Journal of Biological Chemistry·Ramil F LatypovDingjiang Liu
Jul 17, 2012·Analytical and Bioanalytical Chemistry·Dilip ShresthaAttila Jenei
Aug 6, 2013·Biosensors & Bioelectronics·Asta Makaraviciute, Almira Ramanaviciene
Apr 9, 2014·Analytical Chemistry·Shangquan WuQingchuan Zhang
Feb 18, 2015·Journal of Controlled Release : Official Journal of the Controlled Release Society·Yun WuIan C Davis

❮ Previous
Next ❯

Citations

May 29, 2018·Chemical Communications : Chem Comm·Rafael AlonsoÁngel Maquieira
Jan 25, 2021·International Journal of Pharmaceutics·Fatma Betul ArslanSema Calis
Sep 13, 2020·Journal of Nuclear Medicine : Official Publication, Society of Nuclear Medicine·Kyung-Ho JungKyung-Han Lee
Dec 12, 2018·International Journal of Biological Macromolecules·Jyoti GuptaM Saleemuddin
May 15, 2020·Nano Letters·Michael J PoellmannSeungpyo Hong

❮ 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.