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The crude product was purified by a silica gel column chromatography using hexane and dichloromethane mixture (10:1 v/v) as eluent and obtained as yellow powder in 74% yield

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The peptide was first extracted from calf thymus in the 1970s and in the decades since has been subject to a number of studies including human clinical trials

They constructed six progressive deletion mutants within the extra sequence region with altered autoprocessing capabilities and variable catalytic activities (Suzuki and Kumagai, 2002