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Grade:
Vaccine
------------HAVRIX® and VAQTA®
According to the statistics from WHO (1), 80 -95% of the children below 5 years
old do not develop any symptoms after infection, while the risks of developing
clinical symptoms rise with age. Among older children and adults, 75-90% of the
cases will develop symptomatic diseases after infection. Moreover, the mortality
rate is as high as 2.1% for adults above 40 years old. Therefore, vaccination is
developed to provide protection against the diseases.
In the following part, I will summarize the key steps of downstream processes of
Vaqta® and Havrix® respectively, after which an analysis of the two methods
will be discussed.
Seperation of insolubles
Before disrupt the cells with detergent, the bioreactors are drained and the cell
monolayer or microcarriers are rinsed with PBS solution to remove the
remaining culture medium (2). Although the literature did not provide detailed
description of this step, in my perspective, the rinsing step can be viewed as
cake washing which produces a concentrated clean cake of cells.
Cell disruption
The concentrated cells are lysed with detergent Triton X-100, which solubilise
and permeablize the membranes. In the case of microcarrier technology is used
(4), the cell lysate is recycled several times to 5µ m orifices and allow the
microcarriers (MC) and nuclei to settle while the MC free lysate is collected. A
second volume of Triton X-100 is added to the MC beads and recycles through
orifices. The two sets of MC free lysate are pooled together as MC free cell
1
As technology develops, microcarrier fermentation technology is more preferred as it provides larger
surface area for cell growth on the small microbeads(90 to 250 microns in diameter). If microcarrier
technology is used, an additional step of filtering out the microbeads from lysate is needed before filter out
the virus particles from the cell lysate.
lysate preparation. The repeated cell lysis on MC beads helps minimize the
product loss thus improves product yield.
Removal of unwanted cell debris from lysate
The MC free lysate preparation consists of HAV and unwanted contaminants
such as cell debris, proteins, nucleic acids etc. The preparation is firstly
subjected to 0.2µ m filtration to remove the small debris (5). Subsequently, the
filter lysate is treated with nuclease, which digested the nucleic acids (5).
For Havrix®, who harvest virus extracellularly, the downstream process is much
simpler. It can be divided into three main stages, namely, concentrating the
HAV harvested supernatant, nuclease and protease treatment, isolation and
purification of HAV product (7).
Bibliography