Antibody-drug conjugate library prepared by scanning insertion of the aldehyde tag into IgG1 constant regions
| العنوان: | Antibody-drug conjugate library prepared by scanning insertion of the aldehyde tag into IgG1 constant regions |
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| المؤلفون: | Penelope M. Drake, Robyn M. Barfield, Yun Cheol Kim, Igor Rupniewski, William E Haskins, Stefanie Bañas, David Rabuka, Betty C.B. Huang |
| المصدر: | mAbs |
| بيانات النشر: | Informa UK Limited, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, Antibody-drug conjugate, Immunoconjugates, formylglycine, Drug Compounding, Immunology, Glycine, Aldehyde tag, bioconjugation, antibody-drug conjugate, 03 medical and health sciences, chemistry.chemical_compound, Protein structure, Peptide Library, Report, Humans, Immunology and Allergy, Computer Simulation, Amino Acid Sequence, FGE, Aldehydes, Binding Sites, formylglycine generating enzyme, Bioconjugation, Chemistry, Combinatorial chemistry, 030104 developmental biology, ADC, Immunoglobulin G, fGly, site-specific, Immunoglobulin Constant Regions, Protein Binding |
| الوصف: | The advantages of site-specific over stochastic bioconjugation technologies include homogeneity of product, minimal perturbation of protein structure/function, and – increasingly – the ability to perform structure activity relationship studies at the conjugate level. When selecting the optimal location for site-specific payload placement, many researchers turn to in silico modeling of protein structure to identify regions predicted to offer solvent-exposed conjugatable sites while conserving protein function. Here, using the aldehyde tag as our site-specific technology platform and human IgG1 antibody as our target protein, we demonstrate the power of taking an unbiased scanning approach instead. Scanning insertion of the human formylglycine generating enzyme (FGE) recognition sequence, LCTPSR, at each of the 436 positions in the light and heavy chain antibody constant regions followed by co-expression with FGE yielded a library of antibodies bearing an aldehyde functional group ready for conjugation. Each of the variants was expressed, purified, and conjugated to a cytotoxic payload using the Hydrazinyl Iso-Pictet-Spengler ligation to generate an antibody-drug conjugate (ADC), which was analyzed in terms of conjugatability (assessed by drug-to-antibody ratio, DAR) and percent aggregate. We searched for insertion sites that could generate manufacturable ADCs, defined as those variants yielding reasonable antibody titers, DARs of ≥ 1.3, and ≥ 95% monomeric species. Through this process, we discovered 58 tag insertion sites that met these metrics, including 14 sites in the light chain, a location that had proved refractory to the placement of manufacturable tag sites using in silico modeling/rational approaches. |
| تدمد: | 1942-0870 1942-0862 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9225c2d78a6e8b260cf57b9695b16653Test https://doi.org/10.1080/19420862.2018.1512327Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....9225c2d78a6e8b260cf57b9695b16653 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19420870 19420862 |
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