The influence of proline isomerization on potency and stability of anti-HIV antibody 10E8.

Autor: Guttman M; Department of Medicinal Chemistry, University of Washington, Seattle, WA, 98195, USA. mguttman@uw.edu., Padte NN; Aaron Diamond AIDS Research Center, Columbia University Vagelos College of Physicians and Surgeons, 630 W 168th St, New York, NY, 10032, USA., Huang Y; Aaron Diamond AIDS Research Center, Columbia University Vagelos College of Physicians and Surgeons, 630 W 168th St, New York, NY, 10032, USA., Yu J; Aaron Diamond AIDS Research Center, Columbia University Vagelos College of Physicians and Surgeons, 630 W 168th St, New York, NY, 10032, USA., Rocklin GJ; Department of Pharmacology, Northwestern University, Chicago, IL, 60611, USA., Weitzner BD; Department of Biochemistry, University of Washington, Seattle, WA, 98195, USA.; Institute for Protein Design, University of Washington, Seattle, WA, USA., Scian M; Department of Medicinal Chemistry, University of Washington, Seattle, WA, 98195, USA., Ho DD; Aaron Diamond AIDS Research Center, Columbia University Vagelos College of Physicians and Surgeons, 630 W 168th St, New York, NY, 10032, USA., Lee KK; Department of Medicinal Chemistry, University of Washington, Seattle, WA, 98195, USA.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2020 Aug 31; Vol. 10 (1), pp. 14313. Date of Electronic Publication: 2020 Aug 31.
DOI: 10.1038/s41598-020-71184-7
Abstrakt: Monoclonal antibody (mAb) 10E8 recognizes a highly conserved epitope on HIV and is capable of neutralizing > 95% of circulating viral isolates making it one of the most promising Abs against HIV. Solution instability and biochemical heterogeneity of 10E8 has hampered its development for clinical use. We identify the source of 10E8 heterogeneity being linked to cis/trans isomerization at two prolines within the YPP motif in the CRD3 loop that exists as two predominant conformers that interconvert on a slow timescale. The Y trans P conformation conformer can bind the HIV gp41 epitope, while the Y cis P is not binding competent and shows a higher aggregation propensity. The high barrier of isomerization and propensity to adopt non-binding competent proline conformers provides novel insight into the slow binding kinetics, low potency, and poor solubility of 10E8. This study highlights how proline isomerization should be considered a critical quality attribute for biotherapeutics with paratopes containing potential cis proline amide bonds.
Databáze: MEDLINE
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