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Expression and purification of pneumococcal surface protein a of clade 4 in Escherichia coli using hydroxylapatite

Hualong Xi1, Jinfei Yu1, Qing Sun1

  • 1National Engineering Laboratory for AIDS Vaccine, School of Life Science, Jilin University, Changchun, 130012, China.

Protein Expression and Purification
|June 17, 2018
PubMed

Insights

A new method efficiently produces and purifies Streptococcus pneumoniae surface protein A (PspA) from E. coli. This advance offers a promising strategy for developing PspA-based vaccines against pneumococcal diseases.

Area of Science:

  • Microbiology
  • Vaccine Development
  • Protein Expression

Background:

  • * Streptococcus pneumoniae causes severe diseases globally, particularly in vulnerable populations.
  • * Pneumococcal surface protein A (PspA) is a key target for next-generation vaccines.
  • * Current vaccines face limitations, driving the need for alternative strategies.

Purpose of the Study:

  • * To develop an efficient method for producing and purifying PspA clade 4.
  • * To establish a scalable process for recombinant PspA production.
  • * To validate the method's applicability to other PspA clades.

Main Methods:

  • * Expression of soluble PspA4 in an Escherichia coli system.
  • * Purification using hydroxylapatite and ion-exchange chromatography.
  • * Characterization of purified PspA4 using circular dichroism and mass spectrometry.

Main Results:

  • * Achieved high yields of soluble PspA4 (22.8 mg/L) from autoinducing cultures.
  • * Enhanced PspA4 purity from 17% to over 95% in a single chromatographic step.
  • * Demonstrated successful purification of recombinant PspA4 with high yield (43%).

Conclusions:

  • * A simple and efficient method for mass production of PspA4 was established.
  • * The purification technique significantly improves PspA purity.
  • * This method is adaptable for producing PspA proteins from diverse clades, aiding vaccine development.

Keywords:
FermentationHydroxylapatitePneumoniaPspAPurification

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