Serological analysis notes 1-27-13

2015-01-13

azim58 - Serological analysis notes 1-27-13

Concepts to cover
importance of antibodies
-present before symptoms occur
-persistence and stability of antibodies
-antibodies can be used therapeutically such as rituximab and herceptin
-associated with T cells (the development of high-titre IgG requires CD4
T-cell help.)
history before SEREX
-a few antigens at a time with 1D SDS/PAGE and ELISA
first inventor
(a novel strategy using the antibody
repertoire of cancer patients for the molecular definition
of antigens was developed by Pfreundschuh and his
colleagues Sahin and Tu¨ reci [15])
advantages of serex
-can probe for intracellular antigens
-avoid artifacts of cultured cells
variations of serological analysis
-SEREX, phage display, protein microarray, SERPA, ad MAPPing
-SEREX
--limitations of serex
----linear epitopes (-good expression in bacteria, -antigens with the
most transcripts and expression are often detected by SEREX (TAAs of low
abundance are missed by SEREX, -SEREX is time-consuming and
labour-intensive, -antibodies to non-tumor associated antigens)
-microarray
--small sample amount, Proto-Array from Invitrogen with 80,000
recombinant antigens
SERPA
--biased approach (with known analytes) (autoantibody or autoantigen)
--unbiased approach (new analytes can be discovered) (phage display or
cell lysate)
--pro: high throughput and a lot of data, con: cost and a lot of data
-serological proteome analysis (2D electropheresis (of tumor lysate)->
western blotting -> MS, -only linear epitopes can be detected)
--fast, post-translational modifications
--cons (limited material
co-migrating proteins
issues with reproducibility)

SEREX database
(Cancer Immunome Database, contains all the autoantigens identified by
SEREX)
types of antigens identified (including intracellular antigens)
-cancer testis (CT) antigens
-aberrantly expressed antigens such as HER2/neu, p53, and ras
-oncoproteins ( HER-2 / Neu, ras and
c-MYC)
-tumor suppressor proteins p53
-viral antigens
-cell differentiation antigens
-heat shock proteins (This is because HSPs play important roles in cancer
initiation and progression, and are often overexpressed in cancers
[255–257])
^see list of proteins in document

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