Human IgA-binding Peptides Selected from Random Peptide Libraries

Hatanaka, Takaaki; Ohzono, Shinji; Park, Mirae; Sakamoto, Kotaro; Tsukamoto, Shogo; Sugita, Ryohei; Ishitobi, Hiroyuki; Mori, Toshiyuki; Ito, Osamu; Sorajo, K.; Sugimura, Kazuhisa; Ham, Sihyun; Ito, Yuji

doi:10.1074/jbc.m112.389742

Cited by 28 publications

(17 citation statements)

References 31 publications

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“…The first phage display strategy was focused on the discovery of affinity peptides for Fab A (Figure 1a Table S1 identified small molecule affinity ligands for Fabs 22 and comparable to peptide affinity ligands for other applications. 23 These results validated the efficacy of our screening protocol for the identification of peptides specific to a single Fab target.…”

Section: Single Fab Target Phage Displaysupporting

confidence: 63%

Development of phage biopanning strategies to identify affinity peptide ligands for kappa light chain Fab fragments

Nascimento

Mullerpatan

Azevedo

et al. 2019

Biotechnology Progress

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In this work, two phage biopanning strategies were developed to identify affinity peptides for a single Fab and multiple kappa Fabs. For the biopanning rounds, protein L beads were employed to bind Fab targets in a fixed orientation, and NHS functionalized magnetic beads were used to facilitate evaluation of low pH elution conditions. The resulting peptide sequences were synthesized and the binding to different Fabs was evaluated using fluorescence polarization. The first biopanning approach yielded a peptide with similar affinities for two forms of the Fab (recombinantly expressed and post papain‐digestion) as well as the intact antibody. While moderate affinity was observed toward a murine variant of the Fab with the same complementarity determining regions (CDR) region but different framework, minimal binding occurred to a Fab with high sequence homology but containing different CDR loops. The second biopanning strategy yielded a peptide with affinity for all three kappa Fabs indicating that it may be a good lead for the development of more general affinity reagents for recombinant kappa Fabs. Finally, an affinity peptide column was developed, and its efficacy was demonstrated for Fab purification from a complex cell culture fluid mixture. The results presented in this article demonstrate that different peptide‐based phage biopanning strategies can be effectively employed to identify affinity peptide leads for specific Fab and more general kappa Fab purifications.

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Section: Single Fab Target Phage Displaysupporting

confidence: 63%

Development of phage biopanning strategies to identify affinity peptide ligands for kappa light chain Fab fragments

Nascimento

Mullerpatan

Azevedo

et al. 2019

Biotechnology Progress

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show abstract

“…Jacalin, a lectin that binds to the O-linked sugars on the IgA1 hinge, and light chain binding protein-based strategies offer rather limited possibilities. Immobilised bacterial IgA binding proteins, or peptides derived from them, represent a feasible solution [151,152], and IgA-binding peptides selected from random peptide libraries may also have applicability in IgA purification [153].…”

Section: Constraints Of Using Iga Therapeutically and Efforts To Resomentioning

confidence: 99%

IgA: Structure, Function, and Developability

2019

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Immunoglobulin A (IgA) plays a key role in defending mucosal surfaces against attack by infectious microorganisms. Such sites present a major site of susceptibility due to their vast surface area and their constant exposure to ingested and inhaled material. The importance of IgA to effective immune defence is signalled by the fact that more IgA is produced than all the other immunoglobulin classes combined. Indeed, IgA is not just the most prevalent antibody class at mucosal sites, but is also present at significant concentrations in serum. The unique structural features of the IgA heavy chain allow IgA to polymerise, resulting in mainly dimeric forms, along with some higher polymers, in secretions. Both serum IgA, which is principally monomeric, and secretory forms of IgA are capable of neutralising and removing pathogens through a range of mechanisms, including triggering the IgA Fc receptor known as FcαRI or CD89 on phagocytes. The effectiveness of these elimination processes is highlighted by the fact that various pathogens have evolved mechanisms to thwart such IgA-mediated clearance. As the structure–function relationships governing the varied capabilities of this immunoglobulin class come into increasingly clear focus, and means to circumvent any inherent limitations are developed, IgA-based monoclonal antibodies are set to emerge as new and potent options in the therapeutic arena.

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“…4-N,N-dimethylamino-1,8-naphthalimide-bromoacetamide (4-DMN-BA) and 4-N,N-dimethylaminosulfonyl-7-(2-aminoethylamino)-2,1,3-benzoxadiazole-bromoacetamide (DBD-BA) were reacted with T7 phage-displayed randomized peptide library (-SGGG-X3-C-X5-7-C-X3; where X represents any amino acids) [8] [13] independently via the 10BASEd-T as shown in Figure 1, to make fluorogenic libraries with vast diversity (i.e., 10 9 ). As a target protein for the proof-of-concept study, we chose glutathione S-transferase (GST) because we have already evolved tetramethylrhodamine (TMR) and Prodan to GST-specific binders [8] [9].…”

Section: Results and Disussionmentioning

confidence: 99%