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Citation: LIANG Qihui, YANG Yi, SHAO Bing, GAO Ye, SONG Yu, HAN Nanyin. Impact of carrier flow composition on membrane adsorption and aggregation of ovalbumin in asymmetrical flow field-flow fractionation[J]. Chinese Journal of Chromatography, ;2018, 36(5): 480-486. doi: 10.3724/SP.J.1123.2017.12014 shu

Impact of carrier flow composition on membrane adsorption and aggregation of ovalbumin in asymmetrical flow field-flow fractionation

LIANG Qihui ^1,2, ,
YANG Yi ² ,
SHAO Bing ² ,
GAO Ye ¹ ,
SONG Yu ¹ ,
HAN Nanyin ¹

1.
School of Pharmaceutical Sciences, Peking University, Beijing 100191 China;
2.
Beijing Center for Disease Prevention and Control, Beijing 100013 China

Received Date: 13 December 2017

Fund Project: Beijing Natural Science Foundation (No. 7162088).

Asymmetrical flow field-flow fractionation (AF4) is a kind of moderate separation technology for the analysis of macromolecules, including proteins with a wide range of sizes. In the separation channel, the membrane adsorption and aggregation of proteins affected by the carrier fluid (CF) composition lead to changes in analyte recovery and size distribution, thereby restricting the application of AF4 to biomolecules. Different pH levels (6.2, 7.4, 8.2), several types of cations (Na⁺, K⁺, Mg²⁺) and various ion strengths (0-0.1 mol/L)were studied to demonstrate the influence of carrier fluid composition on the membrane adsorption and aggregation of proteins. The results revealed the following:a) higher ion strength of CF resulted in a greater degree of membrane adsorption and aggregation; b) the zeta potential, determined by the pI of the protein and the pH of the CF, influenced the adsorption and aggregation; c) divalent cations (Mg²⁺) caused serious adsorption and aggregation. The experimental results can help us achieve better recovery and mitigate aggregate formation by using the optimal CF components in future AF4 studies. Moreover, the findings indicate that AF4 would find extensive application in protein biochemistry assays.
- asymmetrical flow field-flow fractionation (AF4),
- protein aggregation,
- carrier fluid,
- ion strength,
- protein
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