MRM based validation of target antigen expression in Pituitary Adenoma patients
Evaluation of autoantibody signatures in Pituitary Adenoma patients using human proteome arrays
- Organism: Homo sapiens
- Instrument: TSQ Altis
MRM, Pituitary Adenoma, Antibody, HuProt array
Lab head: Sanjeeva Srivastava
Submitter: Arghya Banerjee
To identify the specific diagnostic biomarkers related to Pituitary Adenomas (PAs), serological antibody profiling was performed for three types of PAs, namely Acromegaly, Cushing’s and Nonfunctional Pituitary Adenomas (NFPAs), using the HuProt™ (Human Proteome Microarray). Autoantibody profile of PAs led to the identification of differentially expressed significant proteins and some of the significant proteins were taken for MRM based validation. Current study describes the autoantibody screening in PAs and identifies the unique autoantibody signatures in Acromegaly, Cushing’s and NFPA
This data was acquired for six proteins; three from Acromegaly vs Control comparison, two proteins from NFPA vs Control and one protein from Cushing vs Control comparison. The transition list was prepared using Skyline Daily software (version 21.2.0), considering peptides between 8 to 25 amino acid length. For the target list of peptides, all y product ions with a +1 charge of +2 precursors were selected. The transition list was utilized for building the MRM based method in TSQ Triple Quadrupole Altis Mass spectrometer (Thermo) connected to Ultimate 3000 high-performance liquid chromatography system. The samples were monitored over an LC gradient of 10 minutes. Two solvent systems included 0.1% Formic acid (FA) in Milli-Q water as solvent A and 0.1% FA in Acetonitrile as solvent B. .
Five individual samples from each group were taken and 1.2 micrograms of the pooled peptide of each group were run against the target list of peptides. The raw data obtained were analyzed in Skyline Daily software, where peaks were manually annotated to mark the correct peak area. Samples were compared for their peak intensity and area between different comparisons as Acromegaly/ Cushing vs Control.
Created on 7/7/22, 1:18 PM