Adipokinetic hormones (AKHs) are insect anti-stress hormones that maintain the biochemical and physiological homeostasis of the insect body (Kodrík, 2008). AKHs are octa, nona- or decapeptides with both termini blocked: the N-terminus by a pyroglutamate residue and the C-terminus by an amide. Typically, a specific antibody and ELISA method are used for their quantification in the insect central nervous system and haemolymph. For the latter, prior to executing the ELISA test, several pre-purification steps are required; they also involve a solid phase extraction cartridge.
ABSTRACT
Prepurification of adipokinetic hormones (AKHs) from insect haemolymph prior to ELISA quantification, using a Micro Spin SpeExtra™ MSPE C18P cartridge (0.22 µm nylon membrane, 15 mg, 0.7 mL). The test peptide is Pyrap-AKH from the firebug Pyrrhocoris apterus (pGlu-Leu-Asn-Phe-Thr-Pro-Asn-Trp-NH2). Recovery is checked by gradient HPLC with fluorescence detection (Ex 280 nm / Em 348 nm), solution A: 0.11% TFA in water, solution B: 0.1% TFA in 60% acetonitrile, flow rate 1.5 mL/min. The MSPE step reaches 97.0% efficiency (relative peak area 265.79 against 273.95 for the control sample). Developed by the Biology Centre CAS, České Budějovice.
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Pyrap-AKH (adipokinetic hormone)CAS number not available
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Biology Centre CAS, České Budějovice
Biology Centre of the Czech Academy of Sciences, České Budějovice (Czech Republic). Named as the developer of the SpeExtra(TM) MSPE application for the pre-purification of insect adipokinetic hormones.
References
Kodrík D. (2008) Adipokinetic hormone functions that are not associated with insect flight. Physiol. Entomol. 33: 171-180.
Kodrík D., Socha R., Šimek P., Zemek R. and Goldsworthy G.J. (2000) A new member of the AKH/RPCH family that stimulates locomotory activity in the firebug, Pyrrhocoris apterus (Heteroptera), Insect Biochem. Mol. Biol. 30: 489-498.
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