Complete antibody digestion with pepsin in just three minutes
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چکیده
Capturem Pepsin was developed for rapid and efficient digestion of antibodies—with a room-temperature protocol (Figure 1)—designed for downstream proteomics analysis. Pepsin is an acidic protease with utility in a wide variety of applications, most notably in antibody sample preparation for characterization by mass spectrometry (MS). The specificity (Keil, B. 1992) and activity of digestion are both pH-dependent, with an active range between pH 1–5. For the purpose of MS, digestions are typically carried out at pH 1–2 in order to cleave peptide bonds at both Nand C-terminal phenylalanine, leucine, tryptophan, and tyrosine residues. Altering the pH of the reaction will, in turn, affect the amount of digestion that occurs; 90% of maximum activity is achieved at pH 1.5, while only 35% activity is achieved at pH 4.5 (Bohak, Z. 1969).
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Improvement of Leishmania major Amastigote ELISA Test Using Various Antigen Antibody Dissociating Agents and Pepsin Digestion.
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