Immunoradiometric assay of carcinoembryonic antigen with use of avidin-biotin labeling.

نویسندگان

  • J Peters
  • H Schmidt-Gayk
  • B Peters
  • F P Armbruster
  • A Quentmeier
  • D Mathias
چکیده

In evaluating an enzyme-linked immunoassay of carcinoembryonic antigen (CEA) we found that the IgG fraction of polyclonal anti-CEA antibodies (DAKO) bound very well to the walls of polypropylene test tubes. We therefore developed an immunoradiometric CEA assay based on this binding of polyclonal anti-CEA antibody. We biotinylated a commercially available monoclonal antibody (Hybritech) and bound this to the CEA-anti-CEA bound to the tube wall. For detection we used 125I-labeled streptavidin. In comparison with several immunoassays for CEA this system offered several advantages such as greater linearity of the standard curve (from 0 to 74 micrograms/L), a steeper dose-response curve, and smaller coefficients of variation in the clinically useful range. This assay system may be used for other large molecules, so that only one tracer, the 125I-labeled streptavidin, has to be labeled; thus the technique seems suited for several different assays.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Immunoradiometric assay of corticotropin with use of avidin-biotin separation.

We have developed a "sandwich"-type immunoradiometric assay for corticotropin (ACTH), with a detection limit of 2 ng/L. Two antibodies are used: a mouse monoclonal antibody directed against ACTH[1-17] and labeled with 125I; and a purified polyclonal goat antibody directed against ACTH[34-39] and conjugated to biotin. We could separate 125I-labeled antibody bound to ACTH from 125I-labeled antibo...

متن کامل

Ultrasensitive, specific, two-antibody immunoradiometric assay that detects free alpha subunits of glycoprotein hormones in blood of nonpregnant humans.

This noncompetitive, sensitive, immunoradiometric assay of the free alpha subunit of human pituitary glycoprotein hormones is based on two monoclonal antibodies and an avidin-biotin separation system. The affinity of the first antibody, mouse anti-alpha subunit covalently conjugated to biotin, is 3.8 x 10(11) L/mol. The second antibody, radiolabeled with 125I, has an affinity of 5.4 x 10(11) L/...

متن کامل

Immunoradiometric assay of human proinsulin and partially processed proinsulin with use of monoclonal antibody and streptavidin-biotin labeling.

The sensitive and specific immunoradiometric assay is described for human proinsulin and its intermediate peptides (65-66 split and 32-33 split proinsulin). We developed a monoclonal antibody-based two-site immunoradiometric assay with use of streptavidin-biotin labeling. The detection limits of the assays lie in the range of 0.5-2.0 pM. In the proinsulin assay proinsulin cross-reacted 66% with...

متن کامل

Preparation of prostate specific antigen standards for immunoradiometric assay

Background: Immunoradiometric assay is one of the most common and precise methods for determination of prostate specific antigen (PSA) in clinical laboratories. Usual use of human serum in routine assays has many disadvantages such as easy contamination and precipitation, instability and unavailability. Thus in order to avoid these problems the artificial matrix was used which acts sim...

متن کامل

Quantification of biotin in serum by competition with solid-phase biotin for binding to peroxidase-avidin conjugate.

We have developed a simple, fast, accurate, and reproducible method--an Enzyme-Linked Ligand Sorbent Assay (ELLSA)--for quantifying biotin without the use of isotopes and immunological reagents. The analytical principle is based on the competition between biotin of the sample and immobilized biotin, i.e., biotinylated bovine serum albumin coated onto immunoplates, for binding to horseradish per...

متن کامل

ذخیره 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 نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Clinical chemistry

دوره 35 4  شماره 

صفحات  -

تاریخ انتشار 1989