[Analyze the bioactivity of PD-1 and PD-L1 recombination protein which expressed by prokaryotic system]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2009 Dec;23(6):424-6.
[Article in Chinese]

Abstract

Objective: To analyze the interaction of PD-1 and PD-L1 recombination protein and to know their bioactivity and affinity.

Methods: Stick the PD-1 protein on the surface of CM5 sensor chip by the method of Ammine coupling after being preconsentrated. Dilute the PD-L1 protein step by step and reject it to the passage on CM5 sensor chip which had been stick by PD-1. The time of combination is 3 minutes and of separation is 15 minutes, respectively. Observe the procession and analyze data by BIA Evaluation software 4.

Results: On the consistency of 40 microg/ml, pH 4.5, the PD-1 protein could couple steady on the surface of CM5sensor chip, RU is 3300. On the density of 200 mmol/ml PD-L1 could combine with PD-1 specifically, RU = 150, K(D) = 3.5 x 10(-6).

Conclusion: The PD-1 and PD-L1 recombination protein which we expressed by prokaryotic system have good affinity and bioactivity. The results could provide basic condition for later study.

MeSH terms

  • Antigens, CD / chemistry
  • Antigens, CD / genetics
  • Antigens, CD / isolation & purification
  • Antigens, CD / metabolism*
  • Apoptosis Regulatory Proteins / chemistry
  • Apoptosis Regulatory Proteins / genetics
  • Apoptosis Regulatory Proteins / isolation & purification
  • Apoptosis Regulatory Proteins / metabolism*
  • B7-H1 Antigen
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Gene Expression*
  • Humans
  • Kinetics
  • Programmed Cell Death 1 Receptor
  • Protein Binding
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism

Substances

  • Antigens, CD
  • Apoptosis Regulatory Proteins
  • B7-H1 Antigen
  • CD274 protein, human
  • PDCD1 protein, human
  • Programmed Cell Death 1 Receptor
  • Recombinant Proteins