Non-affinity purification of a nanobody by void-exclusion anion exchange chromatography and multimodal weak cation exchange chromatography

2019 ◽  
Vol 225 ◽  
pp. 88-96 ◽  
Author(s):  
Xiying Fan ◽  
Yuhang Zhou ◽  
Jianli Yu ◽  
Fei Li ◽  
Quan Chen ◽  
...  
Keyword(s):  
Cation Exchange ◽  
Anion Exchange ◽  
Affinity Purification ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Weak Cation Exchange

scholarly journals CHROMATOGRAPHIC STUDIES OF THE RHEUMATOID FACTOR

1959 ◽  
Vol 110 (2) ◽  
pp. 169-186 ◽  
Author(s):  
J. Lospalluto ◽  
M. Ziff
Keyword(s):  
Cation Exchange ◽  
Rheumatoid Factor ◽  
Anion Exchange ◽  
Gamma Globulin ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Latex Particles ◽  
Factor Ii ◽  
Human Fraction

Serum and serum fractions from patients with rheumatoid arthritis have been subjected to anion exchange chromatography on diethylaminoethyl cellulose. The rheumatoid factor activity was clearly separated from the bulk of the gamma globulin by virtue of the anomalous behavior of macroglobulins on this column. Purification of rheumatoid factor by precipitation with fraction II, resolution in 4 M urea, and two successive fractionations on an anion exchanger yielded a highly active preparation consisting of approximately 95 per cent 19S gamma globulin with no detectable amount of 7S contaminant. Evidence was obtained for the existence of two factors by cation exchange chromatography (carboxymethyl cellulose) of rheumatoid serum and of macroglobulin previously separated by anion exchange chromatography. One factor (factor I) agglutinated sensitized sheep cells and latex particles and also precipitated with human fraction II. The second (factor II) agglutinated latex partides and precipitated with fraction II but did not agglutinate sensitized sheep cells. A substance with the properties of factor II was demonstrated in two of four normal sera by cation exchange chromatography. It was suggested that the agglutination of latex particles and precipitation with human fraction II, observed in the case of certain abnormal sera, may be due to elevated concentrations of factor II.

Characterization of the Acidic Species of a Monoclonal Antibody Using Weak Cation Exchange Chromatography and LC-MS

2015 ◽  
Vol 87 (17) ◽  
pp. 9084-9092 ◽  
Author(s):  
Gomathinayagam Ponniah ◽  
Adriana Kita ◽  
Christine Nowak ◽  
Alyssa Neill ◽  
Yekaterina Kori ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cation Exchange ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Weak Cation Exchange

scholarly journals A Novel Hemagglutinin with Antiproliferative Activity against Tumor Cells from the Hallucinogenic MushroomBoletus speciosus

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Jian Sun ◽  
Tzi-Bun Ng ◽  
Hexiang Wang ◽  
Guoqing Zhang
Keyword(s):  
Cation Exchange ◽  
Antiproliferative Activity ◽  
Gel Filtration ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Lymphocytic Leukemia ◽  
Hemagglutinating Activity ◽  
Cation Exchange Chromatography ◽  
Type Ab

Little was known about bioactive compounds from the hallucinogenic mushroomBoletus speciosus. In the present study, a hemagglutinin (BSH,B. speciosushemagglutinin) was isolated from its fruiting bodies and enzymatic properties were also tested. The chromatographic procedure utilized comprised anion exchange chromatography on Q-Sepharose, cation exchange chromatography on CM-Cellulose, cation exchange chromatography on SP-Sepharose, and gel filtration by FPLC on Superdex 75. The hemagglutinin was a homodimer which was estimated to be approximately 31 kDa in size. The activity of BSH was stable up to 60°C, while there was a precipitous drop in activity when the temperature was elevated to 70°C. BSH retained 25% hemagglutinating activity when exposed to 100 mM NaOH and 25 mM HCl. The activity was potently inhibited by 1.25 mM Hg2+and slightly inhibited by Fe2+, Ca2+, and Pb2+. None of the sugars tested showed inhibition towards BSH. Its hemagglutinating activity towards human erythrocytes type A, type B, and type AB was higher than type O. The hemagglutinin showed antiproliferative activity towards hepatoma Hep G2 cells and mouse lymphocytic leukemia cells (L1210)in vitro, with IC50of 4.7 μM and 7.0 μM, respectively. It also exhibited HIV-1 reverse transcriptase inhibitory activity with an IC50of 7.1 μM.

Novel tetrazole-functionalized ion exchanger for weak cation-exchange chromatography of proteins

2008 ◽  
Vol 1187 (1-2) ◽  
pp. 197-204 ◽  
Author(s):  
Genhu Lei ◽  
Xiaohu Xiong ◽  
Yinmao Wei ◽  
Xiaohui Zheng ◽  
Jianbin Zheng
Keyword(s):  
Cation Exchange ◽  
Exchange Chromatography ◽  
Ion Exchanger ◽  
Cation Exchange Chromatography ◽  
Weak Cation Exchange

Expression and Purification of Tetanus Toxin Fragment C in Escherichia coli BL21(DE3)

2020 ◽  
Vol 27 (11) ◽  
pp. 1132-1140
Author(s):  
Pengdi Chai ◽  
Xiuying Pu ◽  
Jianqiang Li ◽  
Xiaoyu Xia ◽  
Jun Ge ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Molecular Weight ◽  
Secondary Structure ◽  
Cation Exchange ◽  
Anion Exchange ◽  
Tetanus Toxin ◽  
Soluble Proteins ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Simple Method

Background: Tetanus is an infectious disease caused by Clostridium secreting tetanus toxin in anaerobic environment. The fragment C of Tetanus toxin (TTc) has been widely studied as a candidate vaccine to replace the existing tetanus toxoid vaccine. Objective: In this study, we established a simple method to purify recombinant protein TTc with ion-exchange chromatography from Escherichia coli expression systems. Methods: The TTc gene sequence was cloned into pET26b (+) vector and transferred to E. coli BL21 (DE3) for expression. The fermentation conditions (IPTG concentration, Induction temperature, Induction time) were optimized to obtain more soluble proteins. The soluble proteins were purified by Anion exchange chromatography and Cation exchange chromatography. The sequence of columns in the purification process was discussed. Finally, the stability of purified TTc protein were determined, the secondary structure of the purified TTc protein was determined by circular dichroism. The molecular weight of the purified TTc protein was determined by liquid chromatograph- mass spectrometer. Furthermore, we verified the immunogenicity of the purified protein in mice. Results: The purity of TTc improved from 34% to 88% after the first anion exchange column, and the final yield of recombinant TTc (purity > 95%) can reach 84.79% after the following cation exchange chromatography. The recombinant TTc had a molecular weight of 51.737 KDa, was stable at 4 °C and weak alkaline environment, was a β-sheet secondary structure, and had strong immunogenicity. Conclusion: The purification method we developed might be an efficient method for the industrial production of tetanus recombinant TTc vaccine.

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