Effect of acidification on the electrophoretic pattern of anacid gastric juice

Brummer, Pekka; Seppälä, P

doi:10.1007/bf02232029

Cited by 8 publications

(2 citation statements)

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“…In addition, previous studies have shown that acidification of neutralized gastric fluid can reactivate gastric proteases, resulting in autodigestion of proteins. 14,15,55 Therefore, methods that involve acidification of gastric fluid (e.g., TCA precipitation) should be avoided to prevent activation of gastric proteases.…”

Section: ' Discussionmentioning

confidence: 99%

“…These observations underscore the importance of determining gastric fluid pH immediately after its removal from the stomach to ensure proper sample processing and analysis. In addition, previous studies have shown that acidification of neutralized gastric fluid can reactivate gastric proteases, resulting in autodigestion of proteins. ,, Therefore, methods that involve acidification of gastric fluid (e.g., TCA precipitation) should be avoided to prevent activation of gastric proteases.…”

Section: Discussionmentioning

confidence: 99%

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Characterization of the Human Gastric Fluid Proteome Reveals Distinct pH-Dependent Protein Profiles: Implications for Biomarker Studies

Kam

Hennessy²,

Chua

et al. 2011

J. Proteome Res.

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Gastric fluid is a source of gastric cancer biomarkers. However, very little is known about the normal gastric fluid proteome and its biological variations. In this study, we performed a comprehensive analysis of the human gastric fluid proteome using samples obtained from individuals with benign gastric conditions. Gastric fluid proteins were prefractionated using ultracentrifuge filters (3 kDa cutoff) and analyzed by two-dimensional gel electrophoresis (2-DE) and multidimensional LC-MS/MS. Our 2-DE analysis of 170 gastric fluid samples revealed distinct protein profiles for acidic and neutral samples, highlighting pH effects on protein composition. By 2D LC-MS/MS analysis of pooled samples, we identified 284 and 347 proteins in acidic and neutral samples respectively (FDR ≤1%), of which 265 proteins (72.4%) overlapped. However, unlike neutral samples, most proteins in acidic samples were identified from peptides in the filtrate (i.e., <3 kDa). Consistent with this finding, immunoblot analysis of six potential gastric cancer biomarkers rarely detected full-length proteins in acidic samples. These findings have important implications for biomarker studies because a majority of gastric cancer patients have neutral gastric fluid compared to noncancer controls. Consequently, sample stratification, choice of proteomic approaches, and validation strategy can profoundly affect the interpretation of biomarker findings. These observations should help to refine gastric fluid biomarker studies.

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Section: ' Discussionmentioning

confidence: 99%