Yoshihiro Kurano

Establishment of monoclonal antibody against human Apo B-48 and measurement of Apo B-48 in serum by ELISA method

The elevation of chylomicrons and chylomicron remnants in plasma would lead to hyperlipidemia and other complications. Apo B‐48, which is translated and produced in the adult intestine from the same gene as Apo B‐100, is considered to be an essential component of chylomicrons and chylomicron remnants. Using a peptide representing human Apo B‐48 C‐terminal sequence as immunogen, we established a monoclonal antibody, B48‐151, against human Apo B‐48. The specific reactivity for Apo B‐48 of this monoclonal antibody was confirmed using Western blot analysis of human plasma in fractions isolated as chylomicron and VLDL. Then, we developed a simple sandwich ELISA method for the detection of human Apo B‐48 in serum by combining B48‐151 as capturing antibody and HRP‐conjugated‐polyclonal antibodies for Apo B as signaling antibody. The established sandwich ELISA constitutes a simple method to monitor Apo B‐48 level in chylomicrons and chylomicron remnants in human serum. J. Clin. Lab. Anal. 12:289–292, 1998.

Novel rapid immunochromatographic test based on an enzyme immunoassay for detecting nucleocapsid antigen in SARS-associated coronavirus.

A novel severe acute respiratory syndrome‐associated coronavirus (SARS‐CoV) has been discovered. The detection of both antigens and antibodies in SARS‐CoV from human specimens with suspected SARS plays an important role in preventing infection. We developed a novel rapid immunochromatographic test (RICT) based on the sandwich format enzyme immunoassay (EIA) with an all‐in‐one device for detecting the native nucleocapsid antigen (N‐Ag) of SARS‐CoV using monoclonal antibodies (MoAbs), which we produced by immunizing recombinant N‐Ag to mice. RICT is a qualitative assay for respiratory aspirates and serum specimens. With this assay, a positive result can be judged subjectively by the appearance of a blue line on the device 15 min after the sample is applied. RICT with several pairs of MoAbs showed a high sensitivity for the detection of recombinant N‐Ag as well as viral N‐Ag of SARS‐CoV. rSN122 and rSN21‐2 were the best MoAbs for immobilized antibody and enzyme labeling, respectively. With regard to analytical sensitivity, RICT detected N‐Ag at 31 pg/mL for recombinant N‐Ag, and at 1.99×102 TCID50/mL for SARS‐CoV. The specificity of RICT was 100% when 150 human sera and 50 nasopharyngeal aspirates (NSPs) were used. RICT based on an EIA using the rSN122/rSN21‐2 pair is a sensitive, specific, and reliable rapid assay for detecting N‐Ag in SARS‐CoV treated with either heat or Triton X‐100. J. Clin. Lab. Anal. 19:150–159, 2005.