Journal of biological methods, cilt.10, sa.1, 2023 (Hakemli Dergi)
Serum contains proteins that possess important information
about diseases and their progression. Unfortunately, these proteins, which carry
the information in the serum are in low abundance and are masked by other serum
proteins that are in high abundance. Such masking prevents their
identification and quantification. Therefore, removal of high abundance proteins is required
to enrich, identify, and quantify the low abundance proteins. Immunodepletion
methods are often used for this purpose, but there are limitations in
their use because of off-target effects and high costs. Here we presented a robust, reproducible
and cost-effective experimental workflow to remove immunoglobulins and albumin
from serum with high efficiency. The workflow did not suffer from
such limitations and enabled identification of 681 low abundance proteins that were otherwise
undetectable in the serum. The identified low abundance proteins belonged to 21
different protein classes, namely the immunity-related proteins,
modulators of protein-binding activity, and protein-modifying enzymes. They also played roles
in various metabolic events, such as integrin signalling, inflammation-mediated
signalling, and cadherin signalling. The presented workflow can
be adapted to remove abundant proteins from other types of biological material and to
provide considerable enrichment for low-abundance proteins.