Novel fluorescent probe for highly sensitive bioassay using sequential enzyme-linked immunosorbent assay-capillary isoelectric focusing (ELISA-cIEF).

نویسندگان

  • Terence G Henares
  • Yuta Uenoyama
  • Yuto Nogawa
  • Ken Ikegami
  • Daniel Citterio
  • Koji Suzuki
  • Shun-ichi Funano
  • Kenji Sueyoshi
  • Tatsuro Endo
  • Hideaki Hisamoto
چکیده

This paper presents a novel rhodamine diphosphate molecule that allows highly sensitive detection of proteins by employing sequential enzyme-linked immunosorbent assay and capillary isoelectric focusing (ELISA-cIEF). Seven-fold improvement in the immunoassay sensitivity and a 1-2 order of magnitude lower detection limit has been demonstrated by taking advantage of the combination of the enzyme-based signal amplification of ELISA and the concentration of enzyme reaction products by cIEF.

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

“elisa-cief” Using Capillary-based Microdevice: Highly-sensitive Elisa Based on Capillary-isoelectric Focusing of Enzyme Reaction Product

Sensitivity enhancement of enzyme-linked immunosorbent assay (ELISA) in capillary-based microdevice was achieved by the concentration of fluorescent final enzyme reaction product using capillary-isoelectric focusing (CIEF). In order to realize this, fluorescent substrate molecule, which can change its structure into ampholyte ion after an enzyme reaction, was newly synthesized and used. Approxi...

متن کامل

DEVELOPMENT OF A SIMPLE AND SENSITIVE ENZYME- LINKED IMMUNOSORBENT ASSAY (ELISA) FOR CLINICAL MEASUREMENT OF TESTOSTERONE USING PENICILLINASE AS LABEL

An enzyme-linked immunosorbent assay using a homologous combination of antiserum raised against testosterone-3-0-carboxymethyloxime-bovine serum albumin (T-3-0-CMO-BSA ) and penicillinase-labelled T-3-0-CMO was developed. This assay was utilized to measure testosterone in serum samples of male and female subjects. The sensitivity of the assay is 50pg/well and the antibody developed crossrea...

متن کامل

Fluobodies against Bioactive Natural Products and their Application in Fluorescence-Linked Immunosorbent Assay

An enzyme-linked immunosorbent assay (ELISA) using monoclonal antibody (MAb), Fab antibody, and single-chain variable fragment (scFv) antibody has become one of the most promising analytical methods owing to its rapidity, sensitivity, and reliability. Recently, a chimera of green fluorescent protein (GFP) with a scFv antibody, named fluobody, was proposed as a probe for an alternative immunosor...

متن کامل

Capillary isoelectric focusing and high-performance cation-exchange chromatography compared for qualitative and quantitative analysis of hemoglobin variants.

We have developed two assays for complete analysis of hemoglobins (Hbs) in the field of hemoglobinopathies: a high-performance cation-exchange liquid chromatography (HPLC) assay on the weak cation-exchanger Poly Cat A and a two-step capillary isoelectric focusing (CIEF) assay on the neutral-coated capillary from Beckman in a narrow pH gradient. The resolution was satisfactory for both HPLC and ...

متن کامل

Capillary isoelectric focusing and affinity capillary electrophoresis approaches for the determination of binding constants for antibodies to the prion protein.

For the development of specific immunological assays, the binding of a specific antibody (Ab) to the target antigen (Ag) has to be relatively strong. In this study, we have utilized affinity capillary electrophoresis (ACE), a form of capillary zone electrophoresis (CZE) to determine the binding constant (Kb) of specific Abs against bovine serum albumin (BSA) and the healthy prion protein (PrPc)...

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:
  • The Analyst

دوره 138 11  شماره 

صفحات  -

تاریخ انتشار 2013