scholarly journals Optimization of the secretory expression of recombinant human C-reactive protein in Pichia pastoris

3 Biotech ◽  
2017 ◽  
Vol 7 (5) ◽  
Author(s):  
Junming Li ◽  
Chengming Sun ◽  
Lei Chen ◽  
Lihui Sun ◽  
Lijun Duan ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
C Reactive Protein ◽  
Secretory Expression ◽  
Reactive Protein

Purification of Recombinant Mouse C-Reactive Protein from Pichia Pastoris GS115 by Nickel Chelating Sepharose Fast-Flow Affinity Chromatography and P-Aminophenyl Phosphoryl Choline Agarose Resin Affinity Chromatography in Tandem

2021 ◽  
Author(s):  
Bin Cheng ◽  
Di Wu ◽  
Ke Wu ◽  
Xiao-Ping Huang ◽  
Jian-Min Lv ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Affinity Chromatography ◽  
Protein Function ◽  
Recombinant Expression ◽  
Expression System ◽  
Post Translational Modification ◽  
C Reactive Protein ◽  
Reactive Protein ◽  
Phosphoryl Choline ◽  
Fast Flow

Abstract C-reactive protein (CRP) is a circulating marker of inflammation yet with ill-defined biological functions. This is partly due to the uncharacterized activities of endogenous CRP in mice, the major animal model used to define protein function. The hurdles for purification and characterization of mouse CRP are its low circulating levels and the lack of specific antibodies. To clear these hurdles, here we developed an efficient expression system by constructing recombinant Pichia pastoris cells for secretion of native conformation mouse CRP. The recombinant expression of mouse CRP in Escherichia coli failed to yield sufficient amount of native protein, reflecting the importance of post-translational modification of glycosylation in aiding proper folding. By contrast, sufficient amount of native mouse CRP was successfully purified from P. pastoris. Preliminary purification was performed by Nickel Chelating Sepharose Fast-Flow affinity chromatography with 6 × His tags attached to the protein. Subsequently, p-Aminophenyl Phosphoryl Choline Agarose resin affinity chromatography was used for tandem purification. The purified mouse CRP showed native pentamer and capabilities of PC binding. Moreover, the 6 × His tag provides a convenient tool for detecting the interactions of mouse CRP with ligands.

908: Preoperative C-Reactive Protein Elevation as a Predictor of Localized Renal Cell Carcinoma

2007 ◽  
Vol 177 (4S) ◽  
pp. 301-301
Author(s):  
Yasumasa Iimura ◽  
Kazutaka Saito ◽  
Minato Yokoyama ◽  
Hitoshi Masuda ◽  
Tsuyoshi Kobayashi ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Renal Cell ◽  
C Reactive Protein ◽  
Reactive Protein ◽  
Localized Renal Cell Carcinoma
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