Towards a universal disulphide stabilised single chain Fv format: importance of interchain disulphide bond location and vL-vH orientation

Protein Engineering, Design & Selection : PEDS
Eve E WeatherillDavid P Humphreys

Abstract

Engineered introduction of interface interchain disulphide bonds is perceived to be a simple method to increase the stability of single chain Fv (scFv). Six disulphide bond locations have been cited within the literature but the potential for the broad use of each has not been examined. Five of these disulphide bond locations were introduced into one scFv in order to compare their relative effects on expression, thermal stability, percent monomer formation and retention of antigen binding. The disulphide bond position vH44-vL100 was observed to enable the most favourable balance of biophysical properties. The vH44-vL100 disulphide bond was introduced into five additional scFv in both vL-vH and vH-vL orientations in order to investigate its general applicability. Data are presented to show the relative influence of scFv sequence, v-region organisation and interchain disulphide bond on expression yield, thermal stability and percent monomer. Introduction of the vH44-vL100 disulphide bond typically resulted in no or little increase in thermal stability and no change in percent monomer but did confer the benefit of permanently fixing monomer:dimer ratios during purification and analysis.

References

Feb 13, 1990·Biochemistry·R GlockshuberA Plückthun
Aug 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·J S HustonR Crea
Oct 21, 1988·Science·R E BirdM Whitlow
Feb 1, 1997·Nature Biotechnology·M J Coloma, S L Morrison
Apr 1, 1997·Protein Science : a Publication of the Protein Society·Z ZhuP Carter
Dec 9, 2000·Protein Engineering·A SchmiedlS Dübel
Dec 17, 2002·Current Opinion in Biotechnology·Surinder K BatraDavid Colcher
Apr 19, 2006·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Jan PachlUNKNOWN Mycograb Invasive Candidiasis Study Group
Dec 19, 2009·Protein Engineering, Design & Selection : PEDS·Kelly Davis OrcuttK Dane Wittrup
Dec 22, 2009·Protein Engineering, Design & Selection : PEDS·Robert MabryMark Snavely
Aug 10, 1994·Applied Optics·D J Jackson

❮ Previous
Next ❯

Citations

Jun 5, 2014·International Journal of Cancer. Journal International Du Cancer·Ming ChengNai-Kong V Cheung
Aug 2, 2014·Immunopharmacology and Immunotoxicology·Safar FarajniaLeila Rahbarnia
Jun 17, 2014·Biochimica Et Biophysica Acta·Matthias J Feige, Johannes Buchner
Oct 1, 2013·Bioconjugate Chemistry·Hanieh KhaliliSteve Brocchini
Feb 6, 2020·International Journal of Molecular Sciences·Stijn De MunterBart Vandekerckhove
Jul 3, 2017·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Tiancen HuSandra W Cowan-Jacob
Jul 22, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Soichiro YamauchiHiroshi Morioka
Apr 18, 2014·The Journal of Biological Chemistry·Yoko Akazawa-OgawaYoshihisa Hagihara
Mar 7, 2020·Acta Crystallographica. Section F, Structural Biology Communications·Alkistis N MitropoulouAnna M Davies
Sep 4, 2020·International Journal of Molecular Sciences·Annamaria SandomenicoMenotti Ruvo
Dec 22, 2020·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Masahito SatoJotaro Suzuki
Oct 16, 2020·International Journal of Molecular Sciences·Manali S SawantPeter M Tessier

❮ 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.