Derivation of splice junction-specific antibodies using a unique hapten targeting strategy and directed evolution.
Derivation of splice junction-specific antibodies using a unique hapten targeting strategy and directed evolution.
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DOI：
10.1016/j.nbt.2022.06.003
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发表时间：
2022-11-25
影响因子：
5.4
通讯作者：
Weiner, Michael P.
中科院分区：
文献类型：
--
作者：
Fuller, Emily P.;O'Neill, Rachel J.;Weiner, Michael P.
Alternative splicing of RNA occurs frequently in eukaryotic cells and can result in multiple protein isoforms that are nearly identical in amino acid sequence, but have unique biological roles. Moreover, the relative abundance of these unique isoforms can be correlative with diseased states and potentially used as biomarkers or therapeutic targets. However, due to high sequence similarities among isoforms, current proteomic methods are incapable of differentiating native protein isoforms derived from most alternative splicing events. Herein, a strategy employing a nonsynonymous, non-native amino acid (nnAA) pseudo-hapten (i.e. an amino acid or amino acid derivative that is different from the native amino acid at a particular position) as a targeting epitope in splice junction-spanning peptides was successful in directed antibody derivation. After isolating nnAA-specific anti-bodies, directed evolution reduced the antibody’s binding dependence on the nnAA pseudo-hapten and improved binding to the native splice junction epitope. The resulting antibodies demonstrated codependent binding affinity to each exon of the splice junction and thus are splice junction- and isoform-specific. Furthermore, epitope scanning demonstrated that positioning of the nnAA pseudo-hapten within a peptide antigen can be exploited to predetermine the isolated antibody’s specificity at, or near, amino acid resolution. Thus, this nnAA targeting strategy has the potential to robustly derive splice junction- and site-specific antibodies that can be used in a wide variety of research endeavors to unambiguously differentiate native protein isoforms.
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影响因子：
2.2
作者：
Holland, Erika G.;Acca, Felicity E.;Belanger, Kristina M.;Bylo, Mary E.;Kay, Brian K.;Weiner, Michael P.;Kiss, Margaret M.
通讯作者：
Kiss, Margaret M.
DOI：
10.4049/jimmunol.1800557
发表时间：
2018-11-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Nelson AM;Carew NT;Smith SM;Milcarek C
通讯作者：
Milcarek C
影响因子：
5.6
作者：
CHOTHIA, C;LESK, AM
通讯作者：
LESK, AM
影响因子：
4.5
作者：
Martinez, Nicole M.;Pan, Qun;Lynch, Kristen W.
通讯作者：
Lynch, Kristen W.
影响因子：
5.4
作者：
Batonick M;Holland EG;Busygina V;Alderman D;Kay BK;Weiner MP;Kiss MM
通讯作者：
Kiss MM