Purification and Characterization of SibaCec

According to the methods in our previous report [11 (link)], the eluted peak of A1 (Fig. 1a) containing antimicrobial activity was pooled, lyophilized, and further purified by RP-HPLC on a Wondasil C₁₈ column (25 × 0.46 cm). The elution was performed using a linear gradient of 0–60 % acetonitrile containing 0.1 % (v/v) trifluoroacetic acid in 0.1 % (v/v) trifluoroacetic acid/water over 70 min. N-terminal sequence of the purified peptide was done by Edman degradation on an Applied Biosystems pulsed liquid-phase sequencer (model ABI 491).Fig. 1

Purification of SibaCec from the salivary gland of S. bannaense and MALDI–TOF MS. a The filtrate of the salivary gland homogenate of S. bannaense was divided by an Inertsil C₄ RP-HPLC column. b The eluted peak of A1 containing antimicrobial activity was further purified by C₁₈ RP-HPLC column. The purified SibaCec is indicated by an arrow. c MALDI-TOF mass spectrometry analysis of the purified SibaCec

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Wu J., Mu L., Zhuang L., Han Y., Liu T., Li J., Yang Y., Yang H, & Wei L. (2015). A cecropin-like antimicrobial peptide with anti-inflammatory activity from the black fly salivary glands. Parasites & Vectors, 8, 561.

Publication 2015

ABI 491

Acetonitrile Antimicrobial Hplc Maldi Mass spectrometry Peptide Salivary gland Trifluoroacetic acid

Corresponding Organization : Soochow University

Other organizations : Kunming Medical University

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Variable analysis

independent variables

Purification method: RP-HPLC using a Wondasil C18 column (25 × 0.46 cm) with a linear gradient of 0–60% acetonitrile containing 0.1% (v/v) trifluoroacetic acid in 0.1% (v/v) trifluoroacetic acid/water over 70 min

dependent variables

Antimicrobial activity of the eluted peak A1 containing the purified peptide

control variables

Not explicitly mentioned

controls

No positive or negative controls are explicitly mentioned

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