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