US 12,454,548 B2
Method for producing refolded protein by using flow microreactor, and protein refolding apparatus
Daisuke Ejima, Kawasaki (JP); Yuichi Nakahara, Kawasaki (JP); Keisuke Kato, Kawasaki (JP); and Masayo Date, Kawasaki (JP)
Assigned to AJINOMOTO CO., INC., Tokyo (JP)
Filed by AJINOMOTO CO., INC., Tokyo (JP)
Filed on May 5, 2021, as Appl. No. 17/308,594.
Application 17/308,594 is a continuation of application No. PCT/JP2019/043284, filed on Nov. 5, 2019.
Claims priority of application No. 2018-208214 (JP), filed on Nov. 5, 2018.
Prior Publication US 2021/0253629 A1, Aug. 19, 2021
Int. Cl. C07K 1/113 (2006.01); B01J 19/24 (2006.01)
CPC C07K 1/1136 (2013.01) [B01J 19/245 (2013.01)] 5 Claims
OG exemplary drawing
 
1. A protein refolding apparatus, comprising:
a first flow path through which a buffer circulates;
a second flow path through which a solubilized solution of a protein which has become insoluble or lost a higher-order structure thereof circulates; and
a V-shaped micromixer having an inner diameter Y of 0.25 to 1.0 mm in which the first flow path and the second flow path are combined and mixed,
wherein the micromixer is configured for bringing buffer having a flow rate of X1 mL/min into contact with the solubilized solution having a flow rate of X2 mL/min,
a relationship between X1 and Y is shown by the following formula (1), and
a relationship between X1 and X2 is shown by the following formula (2):
Y≤ 1/180X1+⅘  (1)
where 20≤X1≤300, and
X2≤ 1/10X0  (2), and
wherein the micromixer is configured for providing a mixing time of from 10 microseconds to 1,000 microseconds.