Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression

For antibody discovery and engineering, yeast surface display (YSD) of antigen-binding fragments (Fabs) and coupled fluorescence activated cell sorting (FACS) provide intact paratopic conformations and quantitative analysis at the monoclonal level, and thus holding great promises for numerous applic...
Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Meng Mei [verfasserIn] Junhong Li [verfasserIn] Shengchen Wang [verfasserIn] Ki Baek Lee [verfasserIn] Brent L. Iverson [verfasserIn] Guimin Zhang [verfasserIn] Xin Ge [verfasserIn] Li Yi [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2019

Schlagwörter:	yeast surface display Fab divergent promotor ER retention sequence molecular chaperone

Übergeordnetes Werk:	In: Frontiers in Bioengineering and Biotechnology - Frontiers Media S.A., 2014, 7(2019)
Übergeordnetes Werk:	volume:7 ; year:2019

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3389/fbioe.2019.00362

Katalog-ID:	DOAJ044601972

Internformat


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520			\|a For antibody discovery and engineering, yeast surface display (YSD) of antigen-binding fragments (Fabs) and coupled fluorescence activated cell sorting (FACS) provide intact paratopic conformations and quantitative analysis at the monoclonal level, and thus holding great promises for numerous applications. Using anti-TNFα mAbs Infliximab, Adalimumab, and its variants as model Fabs, this study systematically characterized complementary approaches for the optimization of Fab YSD. Results suggested that by using divergent promoter GAL1-GAL10 and endoplasmic reticulum (ER) signal peptides for co-expression of light chain and heavy chain-Aga2 fusion, assembled Fabs were functionally displayed on yeast cell surface with sigmoidal binding responses toward TNFα. Co-expression of a Hsp70 family molecular chaperone Kar2p and/or protein-disulfide isomerase (Pdi1p) significantly improved efficiency of functional display (defined as the ratio of cells displaying functional Fab over cells displaying assembled Fab). Moreover, fusing ER retention sequences (ERSs) with light chain also enhanced Fab display quality at the expense of display quantity, and the degree of improvements was correlated with the strength of ERSs and was more significant for Infliximab than Adalimumab. The feasibility of affinity maturation was further demonstrated by isolating a high affinity Fab clone from 1:103 or 1:105 spiked libraries.
650		4	\|a yeast surface display
650		4	\|a Fab
650		4	\|a divergent promotor
650		4	\|a ER retention sequence
650		4	\|a molecular chaperone
653		0	\|a Biotechnology
700	0		\|a Junhong Li \|e verfasserin \|4 aut
700	0		\|a Shengchen Wang \|e verfasserin \|4 aut
700	0		\|a Ki Baek Lee \|e verfasserin \|4 aut
700	0		\|a Brent L. Iverson \|e verfasserin \|4 aut
700	0		\|a Guimin Zhang \|e verfasserin \|4 aut
700	0		\|a Xin Ge \|e verfasserin \|4 aut
700	0		\|a Li Yi \|e verfasserin \|4 aut
773	0	8	\|i In \|t Frontiers in Bioengineering and Biotechnology \|d Frontiers Media S.A., 2014 \|g 7(2019) \|w (DE-627)74950403X \|w (DE-600)2719493-0 \|x 22964185 \|7 nnns
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912			\|a GBV_USEFLAG_A
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912			\|a SSG-OLC-PHA
912			\|a GBV_ILN_11
912			\|a GBV_ILN_20
912			\|a GBV_ILN_22
912			\|a GBV_ILN_23
912			\|a GBV_ILN_24
912			\|a GBV_ILN_39
912			\|a GBV_ILN_40
912			\|a GBV_ILN_62
912			\|a GBV_ILN_63
912			\|a GBV_ILN_65
912			\|a GBV_ILN_69
912			\|a GBV_ILN_70
912			\|a GBV_ILN_73
912			\|a GBV_ILN_74
912			\|a GBV_ILN_95
912			\|a GBV_ILN_105
912			\|a GBV_ILN_110
912			\|a GBV_ILN_151
912			\|a GBV_ILN_161
912			\|a GBV_ILN_170
912			\|a GBV_ILN_213
912			\|a GBV_ILN_230
912			\|a GBV_ILN_285
912			\|a GBV_ILN_293
912			\|a GBV_ILN_602
912			\|a GBV_ILN_2003
912			\|a GBV_ILN_2014
912			\|a GBV_ILN_4012
912			\|a GBV_ILN_4037
912			\|a GBV_ILN_4112
912			\|a GBV_ILN_4125
912			\|a GBV_ILN_4126
912			\|a GBV_ILN_4249
912			\|a GBV_ILN_4305
912			\|a GBV_ILN_4306
912			\|a GBV_ILN_4307
912			\|a GBV_ILN_4313
912			\|a GBV_ILN_4322
912			\|a GBV_ILN_4323
912			\|a GBV_ILN_4324
912			\|a GBV_ILN_4325
912			\|a GBV_ILN_4338
912			\|a GBV_ILN_4367
912			\|a GBV_ILN_4700
951			\|a AR
952			\|d 7 \|j 2019

Indexfelder

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matchkey_str	article:22964185:2019----::rmtnfbessraeipaefcecbertninnmlc
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callnumber-subject-code	TP
publishDate	2019
allfields	10.3389/fbioe.2019.00362 doi (DE-627)DOAJ044601972 (DE-599)DOAJ6175a1fe8a844265bb39d6a010c182ba DE-627 ger DE-627 rakwb eng TP248.13-248.65 Meng Mei verfasserin aut Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier For antibody discovery and engineering, yeast surface display (YSD) of antigen-binding fragments (Fabs) and coupled fluorescence activated cell sorting (FACS) provide intact paratopic conformations and quantitative analysis at the monoclonal level, and thus holding great promises for numerous applications. Using anti-TNFα mAbs Infliximab, Adalimumab, and its variants as model Fabs, this study systematically characterized complementary approaches for the optimization of Fab YSD. Results suggested that by using divergent promoter GAL1-GAL10 and endoplasmic reticulum (ER) signal peptides for co-expression of light chain and heavy chain-Aga2 fusion, assembled Fabs were functionally displayed on yeast cell surface with sigmoidal binding responses toward TNFα. Co-expression of a Hsp70 family molecular chaperone Kar2p and/or protein-disulfide isomerase (Pdi1p) significantly improved efficiency of functional display (defined as the ratio of cells displaying functional Fab over cells displaying assembled Fab). Moreover, fusing ER retention sequences (ERSs) with light chain also enhanced Fab display quality at the expense of display quantity, and the degree of improvements was correlated with the strength of ERSs and was more significant for Infliximab than Adalimumab. The feasibility of affinity maturation was further demonstrated by isolating a high affinity Fab clone from 1:103 or 1:105 spiked libraries. yeast surface display Fab divergent promotor ER retention sequence molecular chaperone Biotechnology Junhong Li verfasserin aut Shengchen Wang verfasserin aut Ki Baek Lee verfasserin aut Brent L. Iverson verfasserin aut Guimin Zhang verfasserin aut Xin Ge verfasserin aut Li Yi verfasserin aut In Frontiers in Bioengineering and Biotechnology Frontiers Media S.A., 2014 7(2019) (DE-627)74950403X (DE-600)2719493-0 22964185 nnns volume:7 year:2019 https://doi.org/10.3389/fbioe.2019.00362 kostenfrei https://doaj.org/article/6175a1fe8a844265bb39d6a010c182ba kostenfrei https://www.frontiersin.org/article/10.3389/fbioe.2019.00362/full kostenfrei https://doaj.org/toc/2296-4185 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 7 2019
spelling	10.3389/fbioe.2019.00362 doi (DE-627)DOAJ044601972 (DE-599)DOAJ6175a1fe8a844265bb39d6a010c182ba DE-627 ger DE-627 rakwb eng TP248.13-248.65 Meng Mei verfasserin aut Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier For antibody discovery and engineering, yeast surface display (YSD) of antigen-binding fragments (Fabs) and coupled fluorescence activated cell sorting (FACS) provide intact paratopic conformations and quantitative analysis at the monoclonal level, and thus holding great promises for numerous applications. Using anti-TNFα mAbs Infliximab, Adalimumab, and its variants as model Fabs, this study systematically characterized complementary approaches for the optimization of Fab YSD. Results suggested that by using divergent promoter GAL1-GAL10 and endoplasmic reticulum (ER) signal peptides for co-expression of light chain and heavy chain-Aga2 fusion, assembled Fabs were functionally displayed on yeast cell surface with sigmoidal binding responses toward TNFα. Co-expression of a Hsp70 family molecular chaperone Kar2p and/or protein-disulfide isomerase (Pdi1p) significantly improved efficiency of functional display (defined as the ratio of cells displaying functional Fab over cells displaying assembled Fab). Moreover, fusing ER retention sequences (ERSs) with light chain also enhanced Fab display quality at the expense of display quantity, and the degree of improvements was correlated with the strength of ERSs and was more significant for Infliximab than Adalimumab. The feasibility of affinity maturation was further demonstrated by isolating a high affinity Fab clone from 1:103 or 1:105 spiked libraries. yeast surface display Fab divergent promotor ER retention sequence molecular chaperone Biotechnology Junhong Li verfasserin aut Shengchen Wang verfasserin aut Ki Baek Lee verfasserin aut Brent L. Iverson verfasserin aut Guimin Zhang verfasserin aut Xin Ge verfasserin aut Li Yi verfasserin aut In Frontiers in Bioengineering and Biotechnology Frontiers Media S.A., 2014 7(2019) (DE-627)74950403X (DE-600)2719493-0 22964185 nnns volume:7 year:2019 https://doi.org/10.3389/fbioe.2019.00362 kostenfrei https://doaj.org/article/6175a1fe8a844265bb39d6a010c182ba kostenfrei https://www.frontiersin.org/article/10.3389/fbioe.2019.00362/full kostenfrei https://doaj.org/toc/2296-4185 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 7 2019
allfields_unstemmed	10.3389/fbioe.2019.00362 doi (DE-627)DOAJ044601972 (DE-599)DOAJ6175a1fe8a844265bb39d6a010c182ba DE-627 ger DE-627 rakwb eng TP248.13-248.65 Meng Mei verfasserin aut Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier For antibody discovery and engineering, yeast surface display (YSD) of antigen-binding fragments (Fabs) and coupled fluorescence activated cell sorting (FACS) provide intact paratopic conformations and quantitative analysis at the monoclonal level, and thus holding great promises for numerous applications. Using anti-TNFα mAbs Infliximab, Adalimumab, and its variants as model Fabs, this study systematically characterized complementary approaches for the optimization of Fab YSD. Results suggested that by using divergent promoter GAL1-GAL10 and endoplasmic reticulum (ER) signal peptides for co-expression of light chain and heavy chain-Aga2 fusion, assembled Fabs were functionally displayed on yeast cell surface with sigmoidal binding responses toward TNFα. Co-expression of a Hsp70 family molecular chaperone Kar2p and/or protein-disulfide isomerase (Pdi1p) significantly improved efficiency of functional display (defined as the ratio of cells displaying functional Fab over cells displaying assembled Fab). Moreover, fusing ER retention sequences (ERSs) with light chain also enhanced Fab display quality at the expense of display quantity, and the degree of improvements was correlated with the strength of ERSs and was more significant for Infliximab than Adalimumab. The feasibility of affinity maturation was further demonstrated by isolating a high affinity Fab clone from 1:103 or 1:105 spiked libraries. yeast surface display Fab divergent promotor ER retention sequence molecular chaperone Biotechnology Junhong Li verfasserin aut Shengchen Wang verfasserin aut Ki Baek Lee verfasserin aut Brent L. Iverson verfasserin aut Guimin Zhang verfasserin aut Xin Ge verfasserin aut Li Yi verfasserin aut In Frontiers in Bioengineering and Biotechnology Frontiers Media S.A., 2014 7(2019) (DE-627)74950403X (DE-600)2719493-0 22964185 nnns volume:7 year:2019 https://doi.org/10.3389/fbioe.2019.00362 kostenfrei https://doaj.org/article/6175a1fe8a844265bb39d6a010c182ba kostenfrei https://www.frontiersin.org/article/10.3389/fbioe.2019.00362/full kostenfrei https://doaj.org/toc/2296-4185 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 7 2019
allfieldsGer	10.3389/fbioe.2019.00362 doi (DE-627)DOAJ044601972 (DE-599)DOAJ6175a1fe8a844265bb39d6a010c182ba DE-627 ger DE-627 rakwb eng TP248.13-248.65 Meng Mei verfasserin aut Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier For antibody discovery and engineering, yeast surface display (YSD) of antigen-binding fragments (Fabs) and coupled fluorescence activated cell sorting (FACS) provide intact paratopic conformations and quantitative analysis at the monoclonal level, and thus holding great promises for numerous applications. Using anti-TNFα mAbs Infliximab, Adalimumab, and its variants as model Fabs, this study systematically characterized complementary approaches for the optimization of Fab YSD. Results suggested that by using divergent promoter GAL1-GAL10 and endoplasmic reticulum (ER) signal peptides for co-expression of light chain and heavy chain-Aga2 fusion, assembled Fabs were functionally displayed on yeast cell surface with sigmoidal binding responses toward TNFα. Co-expression of a Hsp70 family molecular chaperone Kar2p and/or protein-disulfide isomerase (Pdi1p) significantly improved efficiency of functional display (defined as the ratio of cells displaying functional Fab over cells displaying assembled Fab). Moreover, fusing ER retention sequences (ERSs) with light chain also enhanced Fab display quality at the expense of display quantity, and the degree of improvements was correlated with the strength of ERSs and was more significant for Infliximab than Adalimumab. The feasibility of affinity maturation was further demonstrated by isolating a high affinity Fab clone from 1:103 or 1:105 spiked libraries. yeast surface display Fab divergent promotor ER retention sequence molecular chaperone Biotechnology Junhong Li verfasserin aut Shengchen Wang verfasserin aut Ki Baek Lee verfasserin aut Brent L. Iverson verfasserin aut Guimin Zhang verfasserin aut Xin Ge verfasserin aut Li Yi verfasserin aut In Frontiers in Bioengineering and Biotechnology Frontiers Media S.A., 2014 7(2019) (DE-627)74950403X (DE-600)2719493-0 22964185 nnns volume:7 year:2019 https://doi.org/10.3389/fbioe.2019.00362 kostenfrei https://doaj.org/article/6175a1fe8a844265bb39d6a010c182ba kostenfrei https://www.frontiersin.org/article/10.3389/fbioe.2019.00362/full kostenfrei https://doaj.org/toc/2296-4185 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 7 2019
allfieldsSound	10.3389/fbioe.2019.00362 doi (DE-627)DOAJ044601972 (DE-599)DOAJ6175a1fe8a844265bb39d6a010c182ba DE-627 ger DE-627 rakwb eng TP248.13-248.65 Meng Mei verfasserin aut Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier For antibody discovery and engineering, yeast surface display (YSD) of antigen-binding fragments (Fabs) and coupled fluorescence activated cell sorting (FACS) provide intact paratopic conformations and quantitative analysis at the monoclonal level, and thus holding great promises for numerous applications. Using anti-TNFα mAbs Infliximab, Adalimumab, and its variants as model Fabs, this study systematically characterized complementary approaches for the optimization of Fab YSD. Results suggested that by using divergent promoter GAL1-GAL10 and endoplasmic reticulum (ER) signal peptides for co-expression of light chain and heavy chain-Aga2 fusion, assembled Fabs were functionally displayed on yeast cell surface with sigmoidal binding responses toward TNFα. Co-expression of a Hsp70 family molecular chaperone Kar2p and/or protein-disulfide isomerase (Pdi1p) significantly improved efficiency of functional display (defined as the ratio of cells displaying functional Fab over cells displaying assembled Fab). Moreover, fusing ER retention sequences (ERSs) with light chain also enhanced Fab display quality at the expense of display quantity, and the degree of improvements was correlated with the strength of ERSs and was more significant for Infliximab than Adalimumab. The feasibility of affinity maturation was further demonstrated by isolating a high affinity Fab clone from 1:103 or 1:105 spiked libraries. yeast surface display Fab divergent promotor ER retention sequence molecular chaperone Biotechnology Junhong Li verfasserin aut Shengchen Wang verfasserin aut Ki Baek Lee verfasserin aut Brent L. Iverson verfasserin aut Guimin Zhang verfasserin aut Xin Ge verfasserin aut Li Yi verfasserin aut In Frontiers in Bioengineering and Biotechnology Frontiers Media S.A., 2014 7(2019) (DE-627)74950403X (DE-600)2719493-0 22964185 nnns volume:7 year:2019 https://doi.org/10.3389/fbioe.2019.00362 kostenfrei https://doaj.org/article/6175a1fe8a844265bb39d6a010c182ba kostenfrei https://www.frontiersin.org/article/10.3389/fbioe.2019.00362/full kostenfrei https://doaj.org/toc/2296-4185 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 7 2019
language	English
source	In Frontiers in Bioengineering and Biotechnology 7(2019) volume:7 year:2019
sourceStr	In Frontiers in Bioengineering and Biotechnology 7(2019) volume:7 year:2019
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	yeast surface display Fab divergent promotor ER retention sequence molecular chaperone Biotechnology
isfreeaccess_bool	true
container_title	Frontiers in Bioengineering and Biotechnology
authorswithroles_txt_mv	Meng Mei @@aut@@ Junhong Li @@aut@@ Shengchen Wang @@aut@@ Ki Baek Lee @@aut@@ Brent L. Iverson @@aut@@ Guimin Zhang @@aut@@ Xin Ge @@aut@@ Li Yi @@aut@@
publishDateDaySort_date	2019-01-01T00:00:00Z
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callnumber-first	T - Technology
author	Meng Mei
spellingShingle	Meng Mei misc TP248.13-248.65 misc yeast surface display misc Fab misc divergent promotor misc ER retention sequence misc molecular chaperone misc Biotechnology Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression
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topic_title	TP248.13-248.65 Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression yeast surface display Fab divergent promotor ER retention sequence molecular chaperone
topic	misc TP248.13-248.65 misc yeast surface display misc Fab misc divergent promotor misc ER retention sequence misc molecular chaperone misc Biotechnology
topic_unstemmed	misc TP248.13-248.65 misc yeast surface display misc Fab misc divergent promotor misc ER retention sequence misc molecular chaperone misc Biotechnology
topic_browse	misc TP248.13-248.65 misc yeast surface display misc Fab misc divergent promotor misc ER retention sequence misc molecular chaperone misc Biotechnology
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title	Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression
ctrlnum	(DE-627)DOAJ044601972 (DE-599)DOAJ6175a1fe8a844265bb39d6a010c182ba
title_full	Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression
author_sort	Meng Mei
journal	Frontiers in Bioengineering and Biotechnology
journalStr	Frontiers in Bioengineering and Biotechnology
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author_browse	Meng Mei Junhong Li Shengchen Wang Ki Baek Lee Brent L. Iverson Guimin Zhang Xin Ge Li Yi
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title_sort	prompting fab yeast surface display efficiency by er retention and molecular chaperon co-expression
callnumber	TP248.13-248.65
title_auth	Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression
abstract	For antibody discovery and engineering, yeast surface display (YSD) of antigen-binding fragments (Fabs) and coupled fluorescence activated cell sorting (FACS) provide intact paratopic conformations and quantitative analysis at the monoclonal level, and thus holding great promises for numerous applications. Using anti-TNFα mAbs Infliximab, Adalimumab, and its variants as model Fabs, this study systematically characterized complementary approaches for the optimization of Fab YSD. Results suggested that by using divergent promoter GAL1-GAL10 and endoplasmic reticulum (ER) signal peptides for co-expression of light chain and heavy chain-Aga2 fusion, assembled Fabs were functionally displayed on yeast cell surface with sigmoidal binding responses toward TNFα. Co-expression of a Hsp70 family molecular chaperone Kar2p and/or protein-disulfide isomerase (Pdi1p) significantly improved efficiency of functional display (defined as the ratio of cells displaying functional Fab over cells displaying assembled Fab). Moreover, fusing ER retention sequences (ERSs) with light chain also enhanced Fab display quality at the expense of display quantity, and the degree of improvements was correlated with the strength of ERSs and was more significant for Infliximab than Adalimumab. The feasibility of affinity maturation was further demonstrated by isolating a high affinity Fab clone from 1:103 or 1:105 spiked libraries.
abstractGer	For antibody discovery and engineering, yeast surface display (YSD) of antigen-binding fragments (Fabs) and coupled fluorescence activated cell sorting (FACS) provide intact paratopic conformations and quantitative analysis at the monoclonal level, and thus holding great promises for numerous applications. Using anti-TNFα mAbs Infliximab, Adalimumab, and its variants as model Fabs, this study systematically characterized complementary approaches for the optimization of Fab YSD. Results suggested that by using divergent promoter GAL1-GAL10 and endoplasmic reticulum (ER) signal peptides for co-expression of light chain and heavy chain-Aga2 fusion, assembled Fabs were functionally displayed on yeast cell surface with sigmoidal binding responses toward TNFα. Co-expression of a Hsp70 family molecular chaperone Kar2p and/or protein-disulfide isomerase (Pdi1p) significantly improved efficiency of functional display (defined as the ratio of cells displaying functional Fab over cells displaying assembled Fab). Moreover, fusing ER retention sequences (ERSs) with light chain also enhanced Fab display quality at the expense of display quantity, and the degree of improvements was correlated with the strength of ERSs and was more significant for Infliximab than Adalimumab. The feasibility of affinity maturation was further demonstrated by isolating a high affinity Fab clone from 1:103 or 1:105 spiked libraries.
abstract_unstemmed	For antibody discovery and engineering, yeast surface display (YSD) of antigen-binding fragments (Fabs) and coupled fluorescence activated cell sorting (FACS) provide intact paratopic conformations and quantitative analysis at the monoclonal level, and thus holding great promises for numerous applications. Using anti-TNFα mAbs Infliximab, Adalimumab, and its variants as model Fabs, this study systematically characterized complementary approaches for the optimization of Fab YSD. Results suggested that by using divergent promoter GAL1-GAL10 and endoplasmic reticulum (ER) signal peptides for co-expression of light chain and heavy chain-Aga2 fusion, assembled Fabs were functionally displayed on yeast cell surface with sigmoidal binding responses toward TNFα. Co-expression of a Hsp70 family molecular chaperone Kar2p and/or protein-disulfide isomerase (Pdi1p) significantly improved efficiency of functional display (defined as the ratio of cells displaying functional Fab over cells displaying assembled Fab). Moreover, fusing ER retention sequences (ERSs) with light chain also enhanced Fab display quality at the expense of display quantity, and the degree of improvements was correlated with the strength of ERSs and was more significant for Infliximab than Adalimumab. The feasibility of affinity maturation was further demonstrated by isolating a high affinity Fab clone from 1:103 or 1:105 spiked libraries.
collection_details	GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700
title_short	Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression
url	https://doi.org/10.3389/fbioe.2019.00362 https://doaj.org/article/6175a1fe8a844265bb39d6a010c182ba https://www.frontiersin.org/article/10.3389/fbioe.2019.00362/full https://doaj.org/toc/2296-4185
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author2	Junhong Li Shengchen Wang Ki Baek Lee Brent L. Iverson Guimin Zhang Xin Ge Li Yi
author2Str	Junhong Li Shengchen Wang Ki Baek Lee Brent L. Iverson Guimin Zhang Xin Ge Li Yi
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doi_str	10.3389/fbioe.2019.00362
callnumber-a	TP248.13-248.65
up_date	2024-07-03T23:45:09.799Z
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Prompting Fab Yeast Surface Display Efficiency by ER Retention and Molecular Chaperon Co-expression

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