1. A purification platform for antibodies and derived fragments using a de novo designed affinity adsorbent
- Author
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João Gonçalves, Mara G. Freire, Ulrich Rothbauer, Filipa Trovão, Ana C. A. Roque, Arménio M. J. B. Barbosa, Manuel J. B. Matos, Ana Sofia Pina, DQ - Departamento de Química, and UCIBIO - Applied Molecular Biosciences Unit
- Subjects
Antibody fragments, synthetic ligands ,biology ,Chemistry ,Capture ,Structural diversity ,Filtration and Separation ,02 engineering and technology ,Antibody purification ,021001 nanoscience & nanotechnology ,Combinatorial chemistry ,Analytical Chemistry ,Biopharmaceutical industry ,Adsorption ,Affinity ,020401 chemical engineering ,biology.protein ,0204 chemical engineering ,Antibody ,0210 nano-technology - Abstract
Fundação para a Ciência e Tecnologia (FCT) -PTDC/BII-BIO/28878/2017; co-financed by ERDF under the PT2020 Partnership Agreement LISBOA-01-0145-FEDER-028878; PD/BD/128251/2016 from MIT-PT PhD Bioengineering Systems to MJBM; SFRH/BPD/112543/2015 to AJMB; Applied Molecular Biosciences Unit – UCIBIO (UIDP/04378/2020 ; UIDB/04378/2020); CICECO-Aveiro Institute of Materials (UIDB/50011/2020; UIDP/50011/2020), financed by national funds from FCT. Antibody technologies are the most representative success-case in the biopharmaceutical industry. Widely available purification technologies fail in providing a dedicated universal purification platform that can accommodate antibodies structural diversity, namely antibodies from non-human sources, as chicken IgY, and antigen-binding fragments. In this work, we took inspiration from natural and engineered antibody-binding ligands, to rationally design affinity adsorbents able to capture full-length antibodies and fragments. The one-pot Petasis and Ugi combinatorial reactions were sequentially employed to rapidly generate a library of putative solid-phase adsorbents. The best performing adsorbent yielded a single-step recovery, under mild conditions, of human and chicken whole antibodies, antigen-binding fragments and engineered single-domain antibodies from different complex feedstocks. Due to its simple preparation, the lead antibody adsorbent finds broad applicability as a universal purification platform to increase the availability of antibody technologies in research and development. authorsversion published
- Published
- 2021
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