Decreased secretion and unfolded protein response up-regulation are correlated with intracellular retention for single-chain antibody variants produced in yeast.
Decreased secretion and unfolded protein response up-regulation are correlated with intracellular retention for single-chain antibody variants produced in yeast.
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DOI：
10.1002/bit.22376
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发表时间：
2009-09-01
影响因子：
3.8
通讯作者：
Robinson, Anne Skaja
中科院分区：
文献类型：
--
作者：
Xu, Ping;Robinson, Anne Skaja
Heterologous protein expression can easily overwhelm a cell's capacity to properly fold protein, initiating the unfolded protein response (UPR), and resulting in a loss of protein expression. In the current model of the unfolded protein response, the chaperone BiP modulates the activation of the UPR due to its interactions with the signaling protein Ire1p and newly synthesized proteins. In this research, 4−4−20 scFv variants were generated by rational design to alter BiP binding to newly synthesized scFv proteins or via directed evolution aimed at improved secretion. Interestingly, the predicted BiP binding ability did not correlate significantly with the unfolded protein response. However, pulse-chase analysis of scFv fate revealed that mutants with a decreased ER residence time were more highly secreted, indicating that improved protein folding was more likely the cause for improved secretion. In fact, decreased secretion correlated with increased binding by BiP, as determined by co-immune precipitation studies. This suggests that the algorithm is not useful for in vivo prediction of variants, and that in vivo screens are more effective for finding variants with improved properties.
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影响因子：
46.9
作者：
Walsh, G
通讯作者：
Walsh, G
DOI：
10.1128/cdli.10.4.587-595.2003
发表时间：
2003-07-01
期刊：
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY
影响因子：
--
作者：
Deng, XK;Nesbit, LA;Morrow, KJ
通讯作者：
Morrow, KJ
DOI：
10.1093/protein/10.4.435
发表时间：
1997-04-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Nieba, L;Honegger, A;Pluckthun, A
通讯作者：
Pluckthun, A
影响因子：
2.9
作者：
Kauffman, KJ;Pridgen, EM;Robinson, AS
通讯作者：
Robinson, AS
影响因子：
3.8
作者：
Huang, Dagang;Gore, Patrick R.;Shusta, Eric V.
通讯作者：
Shusta, Eric V.