Purification of tryptic peptides for mass spectrometry using polyvinylidene fluoride membrane.
Indian J Biochem Biophys
;
2001 Aug; 38(4): 274-6
Article
in English
| IMSEAR
| ID: sea-26946
ABSTRACT
A simple procedure for the purification of tryptic peptides, prior to mass spectrometric analysis, using polyvinylidene fluoride membrane (PVDF) is described. The sensitivity of mass spectrometric analysis is such that minor impurities in tryptic peptide digests suppress the signal obtained. However, we obtained useful signal, from a sample that did not yield any spectra earlier, by purifying the sample using PVDF membrane. For this, the tryptic peptide digest was first spotted on the membrane which was then air-dried and washed. Further, the membrane was extracted with trifluoroacetic acid (TFA) and acetonitrile and subjected to mass spectrometric analysis. This procedure enabled us to identify a cross-reactive D1 antigen on the neutrophil surface that bound antibodies that targeted 60 kD Ro autoantigen in systemic lupus erythematosus, an autoimmune disorder.
Full text:
Available
Index:
IMSEAR (South-East Asia)
Main subject:
Peptides
/
Polyvinyls
/
Mass Spectrometry
/
Humans
/
Trypsin
/
Membranes, Artificial
/
Neutrophils
Language:
English
Journal:
Indian J Biochem Biophys
Year:
2001
Type:
Article
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