Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving hydrolase
Reexamination Certificate
2007-01-23
2007-01-23
Tate, Christopher R. (Department: 1655)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving hydrolase
C204S464000
Reexamination Certificate
active
10327342
ABSTRACT:
Polypeptides are electroblotted through a digestion membrane to a composite capture membrane that can be directly analyzed using mass spectrometry. The molecular weights of the fragments generated by the digestion membrane are then used to identify the polypeptide from which they originated. The digestion membrane contains an immobilized protease such as trypsin, which cleaves the electroblotted polypeptides into fragments during electroblotting with such high enzyme cleavage capacity and efficiency that one pass of the polypeptide through the membrane is sufficient. The peptide fragments are collected onto a composite capture membrane that is chemically treated, for example by adding a mixture of nitrocellulose and MALDI matrix, so as to absorb peptides near the surface to facilitate desportion, thereby increasing the sensitivity of subsequent analysis by MALDI-TOF mass spectrometry. A wide variety of application are disclosed including identifying proteins separated on a gel or within a tissue sample.
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Huang Yulin
Lotti Robert J.
Nadler Timothy K.
Parker Kenneth G.
Vella George J.
Karnakis Andrew T.
PerSeptive Biosystems Inc.
Tate Christopher R.
Wood Amanda P.
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