In vitro production of recombinant antibody fragments in Pichia pastoris.
In vitro production of recombinant antibody fragments in Pichia pastoris.
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在毕赤酵母中体外生产重组抗体片段。
DOI：
10.1016/s0923-2494(98)80012-6
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Eldin,P
中科院分区：
文献类型：
--
作者：
Pennell,CA;Eldin,P
The methyltropic yeast Pichia pastoris is rapidly becoming a preferred host for the efficient expression of heterologous proteins (reviewed in Hollenberg and Gellissen, 1997; Sreekrishna et al., 1997; Sudbery, 1996). P. pastoris combines the general features of protein expression in eukaryotes with the fast growth and genetic modifiability of prokaryotes. It is less expensive than other eukaryotic expression systems, such as baculovirus or mammalian tissue culture, and it typically yields higher quantities of secreted functional proteins. The ability to correctly express folded secreted proteins, including highly disulphide-bonded ones (White et al., 1994), provides a distinct advantage over bacterial systems that often require laborious and inefficient procedures to denature and refold proteins expressed as insoluble, inclusion bodies (Skerra, 1993). P. pastoris is a superior expression host over its more famous nonmethyltropic relative, Saccharomyces cerevisiae, because P. pastoris grows stably to high cell densities in fermentors (> 100 mg dry weight per ml culture) and it has strong, tightly regulated promoters (Ellis et al., 1985). For these reasons, P. pastoris has recently been exploited as an expression system for the high-level secretion of many proteins, including recombinant antibody fragments. By definition, methyltropic yeasts are capable of utilizing methanol as their sole carbon source. The first enzyme in the methanol-utilization pathway, alcohol oxidase (AOX), is encoded by two closely related genes: AOXI and AOX (Ellis et al., 1985; Cregg et al., 1989). Although the AOXl and AOX proteins have 97% sequence identity and equivalent enzymatic activity, over 95% of the alcohol oxidase activity in P. pastoris is attributable to AOXl. This is due to the strength of the AOXI promoter (Cregg et al., 1989; Koutz et al., 1989). Upon the addition of methanol, AOXI gene transcription is rapidly induced to high levels and ultimately accounts for 5% of the total polyA+ RNA (Cregg and Madden, 1988). The AOXl protein is correspondingly overexpressed and comprises up to 30% of the total intracellular protein. Heterologous genes are therefore cloned under control of the AOXZ promoter to
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血栓调节蛋白小片段的大规模表达、纯化和表征：第六结构域和蛋氨酸 388 的作用。
DOI：
10.1093/protein/8.11.1177
发表时间：
1995
期刊：
Protein engineering
影响因子：
--
作者：
White,CE;Hunter,MJ;Meininger,DP;White,LR;Komives,EA
通讯作者：
Komives,EA
影响因子：
2.7
作者：
Stefan Linder;Manfred Schliwa;E. Kube-Granderath
通讯作者：
Stefan Linder;Manfred Schliwa;E. Kube-Granderath
DOI：
10.1038/nbt0395-255
发表时间：
1995-03-01
期刊：
BIO-TECHNOLOGY
影响因子：
--
作者：
RIDDER, R;SCHMITZ, R;GRAM, H
通讯作者：
GRAM, H
通过专门针对表达 erbB2 原癌基因的细胞的嵌合 B7-2 抗体融合蛋白共同刺激 T 细胞增殖。
影响因子：
4.4
作者：
B. Gerstmayer;U. Altenschmidt;M. Hoffmann;W. Wels
通讯作者：
W. Wels
免疫球蛋白片段的细菌表达。
影响因子：
7
作者：
A. Skerra
通讯作者：
A. Skerra