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SCIENCE CHINA Life Sciences, Volume 59 , Issue 6 : 584-588(2016) https://doi.org/10.1007/s11427-016-5037-2

Quality-control method for the determination of biological activity of engineered calcineurin subunit B

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  • ReceivedDec 21, 2015
  • AcceptedFeb 1, 2016
  • PublishedApr 1, 2016

Abstract


Funded by

National Important Novel Medicine Research Project(2012ZX09304010,2013ZX09102062)

National Natural Science Foundation of China(31270849)


Acknowledgment

Acknowledgements This work was supported by the National Important Novel Medicine Research Project (2012ZX09304010, 2013ZX09102062), and the National Natural Science Foundation of China (31270849).


Open access

This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.


Interest statement

Compliance and ethics The author(s) declare that they have no conflict of interest.


References

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  • Figure 1

    Enzyme kinetics curve relating reaction time to absorption measurements. Note: The kinetics curve represents an example of substrate decomposition by CNA316 promoted by CNB, read from one well of 96-well plate as absorbance versus time. Due to the changing temperature, the initial reaction rate is relatively slow. The middle of the curve approaches a straight line, where the reaction proceeds at the maximum rate (Vmax). As the substrate is consumed, the reaction rate decreases.

  • Figure 2

    Dose-response curve between CNB dilution and Vmax. The S-shaped curve reflects the maximum decomposition rate of pNPP at different concentrations (dilutions) of CNB, where undiluted CNB preparation=1. ΔCNA316 and pNPP concentrations, and the temperature were constant. Two curves in the figure were used for biological activity value referral: ○, represents data for CNB reference material. Δ, represents data for CNB sample.

  • Figure 3

    Analysis of reaction consistency. The maximum reaction rate obtained from the middle segment of the S-shaped curve in Figure 2 shows a linear relationship with logarithmic-transformed dilutions of CNB. The curves were used to analyze parallelism and to assess consistency of CNB samples and CNB reference material. ◇, represent data for CNB reference material. Line equation for data fitting, Y=3.138X+12.129 (R2=0.986). □, represent data for CNB sample. Line equation for data fitting, Y=3.204X+12.017 (R2=0.982).

  • Figure 4

    The effect of CNAΔ316 concentrations on Vmax. Each curve represents the relationship between CNB dilutions and maximum pNPP decomposition rate at the specified CNAΔ316 concentrations. CNAΔ316 concentrations, bottom to top, were: 2.5 μg mL-1, 5 μg mL-1, and 10 μg mL-1.

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