Affinity Switching for Lysozyme and Dual-responsive Microgels by Stopped-flow Technique:Kinetic Control and Activity Evaluation

Yun-zhu Wu; Zhi-huang Zhang; Xin Han; Jian Zhang; Wen-ming Zhang; Jun Yin

doi:10.1007/s10118-017-1948-z

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Affinity Switching for Lysozyme and Dual-responsive Microgels by Stopped-flow Technique:Kinetic Control and Activity Evaluation

Papers | Updated：2021-02-18

- Affinity Switching for Lysozyme and Dual-responsive Microgels by Stopped-flow Technique:Kinetic Control and Activity Evaluation
- Chinese Journal of Polymer Science Vol. 35, Issue 8, Pages: 950-960(2017)
- Affiliations：
  
  Department of Polymer Science and Engineering, School of Chemistry and Chemical Engineering, Hefei University of Technology and Anhui Key Laboratory of Advanced Functional Materials and Devices, Hefei 230009, China
- Author bio：
  
  Jun Yin (殷俊),E-mail:yinjun@hfut.edu.cn
- Funds：
- DOI：10.1007/s10118-017-1948-z
  CLC：
- Received：09 February 2017，
  
  Revised：2017-02-22，
  
  Accepted：07 March 2017，
  
  Published：2017-08
- Accepted：
Scan QR Code
Yun-zhu Wu, Zhi-huang Zhang, Xin Han, et al. Affinity Switching for Lysozyme and Dual-responsive Microgels by Stopped-flow Technique:Kinetic Control and Activity Evaluation[J]. Chinese Journal of Polymer Science, 2017, 35(8): 950-960.
DOI：

Yun-zhu Wu, Zhi-huang Zhang, Xin Han, et al. Affinity Switching for Lysozyme and Dual-responsive Microgels by Stopped-flow Technique:Kinetic Control and Activity Evaluation[J]. Chinese Journal of Polymer Science, 2017, 35(8): 950-960. DOI： 10.1007/s10118-017-1948-z.

摘要

Abstract

The use of proteins as therapeutics in nanomedicine is an emerging research field and has developed rapidly. However

proteins are always vulnerable to renal excretion or digestion by the proteolytic system

in vivo

which limits their usage to a large extent. Although biocompatible polymers have been covalently linked to proteins to protect them from recognition by the immune system and prolong their circulation time

the biological activity of them is sometimes decreased. To fill this gap

physical isolation

wrapping

or encapsulation techniques are employed. Up to now

various mature examples were reported

but the whole time scales for guest molecules loading and releasing

especially the initial rapid loading process

were rarely mentioned. Herein

a series of dual-responsive poly(

-isopropylacrylamide-

-methacrylic acid) (P(NIPAM-

-MAA)) microgels were synthesized and employed to investigate the kinetics of

in situ

complexation and release of lysozyme under external stimuli modulation upon a stopped-flow apparatus

which was suitable for rapid dynamic monitoring. Close inspection of the adsorption kinetics during the early stages (

50 s) revealed that the initial microgel collapse occurred within~1 s

with more rapid transitions being observed when higher lysozyme concentrations were targeted. All the dynamic traces could be well fitted with a double exponential function

suggesting a fast (

) and a slow (

) relaxation time

respectively. Then

the kinetics of releasing bound lysozyme from microgels was carried on by utilizing the pH-responsive property

and the evaluation of the activity of released lysozyme was synchronously measured in a Micrococcus lysodeikticus (

M. lysodeikticus

) cell suspension. The corresponding relaxation time (

) was also calculated by fitting the recorded dynamic traces. We speculate that this work can provide basic dynamics data and theoretical basis for microgels based nanocarriers to be used for protein delivery

controlled release

and possible chemical separation.

关键词

Keywords

references

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