Matches in SemOpenAlex for { <https://semopenalex.org/work/W1437335685> ?p ?o ?g. }
- W1437335685 abstract "KD-247 is a humanized monoclonal antibody that targets the third hypervariable (V3) loop of gp120. It can efficiently neutralize a broad panel of clade B, but not non-clade B, HIV-1 isolates. To overcome this limitation, we are seeking to prepare genetically-engineered single-chain variable fragments (scFvs) of KD-247 that will have broader neutralizing activity against both clade B and non-clade B HIV-1 isolates. Initial attempts of optimizing the expression of KD-247 scFv have resulted in the formation of insoluble protein. Therefore, we have established purification protocols to recover, purify, and refold the KD-247 scFv from inclusion bodies. The protocol involved step-wise refolding of denatured scFv by dilution, dialysis, and on-column nickel-affinity purification. Monomeric scFv was further purified by size-exclusion chromatography. Using far UV circular dichroism (CD) spectroscopy we confirmed the expected beta-sheet profile of the refolded KD-247 scFv. Importantly, the refolded KD-247 scFv showed neutralizing activity against replication-competent HIV-1 BaL and JR-FL Env pseudotyped HIV-1, at potency comparable to that of the native full-size KD-247 antibody. Ongoing studies focus on the application of this system in generating KD-247 scFv variants with the ability to neutralize clade B and non-clade B HIV-1 isolates." @default.
- W1437335685 created "2016-06-24" @default.
- W1437335685 creator A5006337398 @default.
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- W1437335685 date "2012-12-22" @default.
- W1437335685 modified "2023-10-07" @default.
- W1437335685 title "Preparation of biologically active single-chain variable antibody fragments that target the HIV-1 gp120 V3 loop." @default.
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- W1437335685 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3612353" @default.
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- W1437335685 hasPublicationYear "2012" @default.
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