Ultrasound-accelerated immunoassay, as exemplified by enzyme immunoassay of choriogonadotropin.
نویسندگان
چکیده
The rate-limiting step in many solid-phase immunoassays is associated with the slow kinetics of binding of macro-molecular antigen and conjugate to the immobilized phase. We demonstrate that the use of ultrasonic energy to enhance mass transport across liquid/solid interfaces can dramatically accelerate antigen binding to immobilized antibodies. We use an ultrasound-accelerated procedure with an enzyme-channelling test strip containing glucose oxidase and specific antibody to the alpha-subunit of human choriogonadotropin (HCG) co-immobilized onto a cellulose support. A horseradish peroxidase conjugate of monospecific antibody to the beta-subunit of HCG is used in the liquid phase to complete the immune "sandwich." Use of ultrasound to accelerate binding and of enzyme channelling to eliminate wash steps result in a simple two-incubation protocol by which 25 int. units of urinary HCG per liter can be detected visually in less than 20 min of assay time.
منابع مشابه
Evaluation of five kits for detecting choriogonadotropin in urine.
We report an evaluation of the performance characteristics, accuracy, and sensitivity of five test kits designed to detect human choriogonadotropin (hCG) in urine: "Sensitex" (latex agglutination inhibition), "beta-Neocept" (hemagglutination inhibition), "Duoclone" (direct agglutination), "Tandem-Visual HCG" (enzyme immunoassay), and "Sensichrome" (enzyme immunoassay). For comparison, all speci...
متن کاملQuantification of Human Chorionic Gonadotropin by Bovine Serum Albumin Nanoparticles
Background and Aims: Some nanoparticles can be used in immunoassays to increase sensitivity. This study aimed to evaluate a novel nano-immunoassay based on bovine serum albumin nanoparticles (BSA NPs). Materials and methods: At first, the nanostructure was synthesized, and then applied as a tag in the nano-immunoassay. Then the concentration of β-subunit of human chorionic gonadotropin ...
متن کاملComparison of Enzyme Immunoassay, Immunochromatography, and RNA-Polyacrylamide-Gel Electrophoresis for Diagnosis of Rotavirus Infection in Children with Acute Gastroenteritis
Human rotavirus is a major etiologic agent for infantile diarrhea worldwide. It is responsible for up to 3.3 million deaths per year in children in developing countries. Various rapid and sensitive techniques have been developed to readily diagnose rotavirus gastroenteritis. In the present study, we compared the sensitivity and specificity of immunochromatography and RNA-polyacrylamide-gel elec...
متن کاملDETERMINATION OF SERUM CORTISOL LEVEL BY A DIREC T ENZYME IMMUNOASSAY USING PENICILLINASE AS LABEL
Serum cortisol level was measured by an enzyme immunoassay (EIA) using the enzyme penicillinase as a label without prior extraction and purification. Polyclonal antibodies were raised against cortisol-3-ortho-carboxymethyl-oxime (cortisol-3-0-CMO) conjugated to bovine serum albumin (BSA). This antibody showed a very low cross-reactivity with structurally related steroids (3.7% for corticos...
متن کاملSimple enzyme immunoassay for the simultaneous measurement of whole choriogonadotropin molecules and free beta-subunits in sera of women with abnormal pregnancies or tumors of the reproductive system.
A multiple enzyme immunoassay (multi-EIA) was developed to quantify whole-molecule human choriogonadotropin (w-hCG) and free hCG beta-subunits (hCG-beta) simultaneously. A clone of a specific monoclonal antibody was coupled to solid phase; two other clones of different monoclonal antibodies were conjugated to horseradish peroxidase (HRP; EC 1.11.1.7) and alkaline phosphatase (AP; EC 3.1.3.1), r...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Clinical chemistry
دوره 30 9 شماره
صفحات -
تاریخ انتشار 1984