A quantitative immunoassay using chicken antibodies for detection of native and recombinant alpha-amidating enzyme.
A sensitive competitive ELISA has been developed for the detection and quantitation of native and recombinant alpha-amidating enzyme. Chickens immunized with purified enzyme (75 kDa) isolated from a rat medullary thyroid carcinoma, produced IgY antibodies specific for the native enzyme. The assay is defined by a standard curve with a linear range of 0.78-12.5 ng/ml in phosphate-buffered saline, and a limit of sensitivity for detection of the enzyme of 0.20 ng/ml. The immunoassay is capable of detecting enzyme from both tumor derived sources, and from cells genetically engineered to secrete the enzyme into tissue culture medium containing up to 10% fetal calf serum.
Sturmer
A M
AM
Department of Immunology, Unigene Laboratories, Fairfield, NJ 07004.
Driscoll
D P
DP
Jackson-Matthews
D E
DE
D000818:Animals; D000918:Antibody Specificity; D002455:Cell Division; D002478:Cells, Cultured; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007114:Immunization; D007136:Immunoglobulins / Q000096:biosynthesis / Q000302:isolation & purification; D066298:In Vitro Techniques; D007273:Injections, Intramuscular; D007279:Injections, Subcutaneous; D006899:Mixed Function Oxygenases / Q000008:administration & dosage / Q000032:analysis*; D009097:Multienzyme Complexes; D011994:Recombinant Proteins / Q000032:analysis; D012680:Sensitivity and Specificity
C016512:IgY; D007136:Immunoglobulins; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase