A sensitive and specific competition microELISA for the immunoactive polypeptide parathymosin and detection of this peptide in porcine tissues.
A sensitive and specific microELISA assay is described for the immunoactive polypeptide parathymosin. Antibodies against a synthetic peptide corresponding to the rat parathymosin sequence 5-30 were raised in rabbits immunised with this peptide conjugated to keyhole limpet hemocyanin (KLH). The useful range of the assay was 0.25-30 pmol (3-330 ng) of parathymosin and the assay was specific. The related immunoactive polypeptides prothymosin alpha or thymosin alpha 1 showed no cross-reactivity. In spiking experiments the recovery of the assay was found to be greater than 92% at all concentrations tested. The intra-assay variation was 17%, whereas the inter-assay variation was 26%. Using this assay the highest concentration of parathymosin was found in porcine liver, followed by kidney, lung, thymus and spleen. This assay compares favorably with one microELISA and two RIA methods already published, in that it is more sensitive by at least an order of magnitude, and it is simpler and quicker.
Laboratory of Biological Chemistry, University of Ioannina Medical School, Greece.
M
M
Economou
G K
GK
Papadopoulos
K
K
Seferiadis
E P
EP
Heimer
A M
AM
Felix
O
O
Tsolas
D000818:Animals; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000032:analysis; D006433:Hemocyanins; D000922:Immunotoxins; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D008297:Male; D012680:Sensitivity and Specificity; D013154:Spleen / Q000378:metabolism; D013552:Swine; D013947:Thymosin / Q000031:analogs & derivatives* / Q000032:analysis / Q000096:biosynthesis; D013950:Thymus Gland / Q000378:metabolism
D000939:Epitopes; D000922:Immunotoxins; D013947:Thymosin; D006433:Hemocyanins; C044498:parathymosin alpha; C032808:keyhole-limpet hemocyanin