Cytiva eBook - 8

PODCAST

Management of detection and quantification of host cell proteins (HCPs)

A coverage assay compares the total HCP population against HCPs in
a sample, enabling you to determine how many of the proteins are
detected by the ELISA.
The U.S. Pharmacopoeia describes two methods for coverage
assays. One approach involves separating HCPs by 2D electrophoresis
and then performing a Western blot using ELISA antibodies. You
can count the number of total HCPs and the number of the spots
detected by the ELISA antibodies.
The second coverage assay method involves capturing HCPs in a
sample using an affinity column prepared with HCP ELISA antibodies.
The captured proteins are then compared with the original total HCP
population using 2D electrophoresis.
While both methods provide an effective means of determining
ELISA coverage, their reliance on 2D gel electrophoresis-which can
be subject to gel-to-gel variation and other reproducibility issues-
means neither is 100% effective.
To address these challenges, we have introduced the 2D DIGE
multiplex fluorescent prelabeling technology to the coverage assay.
This enables two different samples to coexist on the same
membrane or gel. So there is no need to make any 2D spot pattern
alignment from different 2D gels.
This solves many of the drawbacks of 2D electrophoresis and
provides much accurate and reliable coverage data.
We offer this 2D DIGE/Western blot approach as 2D DIBEā„¢ (differential
in-blot electrophoresis) to our customers. This is all about the methods
biologics developers use to validate the HCP ELISA, but there are also
other important aspects related to the ELISA methodology itself.

8

| GENengnews.com


http://www.GENengnews.com

Cytiva eBook

Table of Contents for the Digital Edition of Cytiva eBook

Contents
Cytiva eBook - 1
Cytiva eBook - 2
Cytiva eBook - 3
Cytiva eBook - Contents
Cytiva eBook - 5
Cytiva eBook - 6
Cytiva eBook - 7
Cytiva eBook - 8
Cytiva eBook - 9
Cytiva eBook - 10
Cytiva eBook - 11
Cytiva eBook - 12
Cytiva eBook - 13
Cytiva eBook - 14
Cytiva eBook - 15
Cytiva eBook - 16
Cytiva eBook - 17
Cytiva eBook - 18
https://www.nxtbookmedia.com