Thesis

Expression and Purification of PiuA Recombinant Protein Using Additives For Protein Carrier in Pneumococcal Conjugate Vaccine Candidate

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Glycoconjugate vaccine involved the conjugation of polysaccharides from a pathogen and a
protein carrier whereas pneumococcal conjugate vaccine (PCV) is one of the examples. The
pneumococcal diseases caused by Streptococcus pneumoniae can be tackled by the PCV, which was
produced by chemical conjugation. Due to the challenges in the production such as polysaccharide
extraction and purification that leads to yield loss, Protein Glycan Coupling Technology (PGCT) offers
a new approach for glycoconjugate vaccine. The recombinant protein PiuA is exhibited to have the
potential as a protein carrier for glycoconjugate vaccine candidates. To improve the expression and
production of PiuA, additives such as SDS and Triton X-100 were utilized. In this project, Triton X-100
with a concentration of 0.1% at 24 hours of cultivation was shown to have the highest expression and
production of PiuA, in which the protein content was 50.9184 µg/mL. The purification process using
affinity chromatography with Ni-NTA resin was also shown to purify the PiuA production, in which
the total PiuA content was 241.8515 µg, and the PiuA content in initial crude was 53.7447 µg/mL.
The percentage purity of PiuA after purification was 98.9%. The recommendations for this project are
to optimize the variety concentrations of the rhamnose, to analyse the rhamnose content during the
cultivation, and a better protein quantification method such as the BCA assay.

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Availability

Detail Information

Series Title

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Call Number

BT 23-002

Publisher

i3L, Jakarta : i3L, Jakarta., 2023

Collation

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Language

English

ISBN/ISSN

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Classification

NONE

Content Type

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Media Type

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Carrier Type

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Edition

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Subject(s)

protein expression
pneumococcal conjugate vaccine (PCV)
recombinant protein
affinity chromatography
additives
PGCT
protein carrier

Specific Detail Info

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Statement of Responsibility

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