BACKGROUND: Apolipoprotein B-48 (apoB-48) is produced by the small intestine, as a part of chylomicrons (CMs), and appears to be a suitable marker for clinical studies of postprandial lipoproteins and related cardiovascular risk factors. We have developed an assay for routine analysis to quantify apoB-48 in serum or plasma. METHODS: A microtiter plate was coated with monoclonal antibody (4C8) raised against apoB-48 C-terminal specific decapeptide. Serum samples were diluted 100-fold with 0.05 mol/l Tris-HCl buffer (with or without 0.1% Triton X-100). Appropriate calibration curves were obtained in the ELISA by using apoB-48 recombinant antigen. RESULTS: No cross-reactivity (<0.001%) was found with apoB-100, as verified by ELISA and Western blot analyses. Intra- and inter-assay CVs were 4.8% and 5.4%, respectively. Recovery of added recombinant apoB-48 in serum was within 94-105%. The assay linearity was intact >5-fold dilution of serum by dilution buffer. ApoB-48 levels in healthy controls (n=18) at fasting were within the range of 2.69-6.56 microg/ml (mean+/-S.D.: 4.60+/-1.54 microg/ml). In healthy subjects, serum apoB-48 concentrations markedly increased in the postprandial state, in parallel with serum triglycerides. CONCLUSION: This method for measuring apoB-48 using the monoclonal antibody 4C8 is simple, reliable and suitable for routine analyses.
BACKGROUND:Apolipoprotein B-48 (apoB-48) is produced by the small intestine, as a part of chylomicrons (CMs), and appears to be a suitable marker for clinical studies of postprandial lipoproteins and related cardiovascular risk factors. We have developed an assay for routine analysis to quantify apoB-48 in serum or plasma. METHODS: A microtiter plate was coated with monoclonal antibody (4C8) raised against apoB-48 C-terminal specific decapeptide. Serum samples were diluted 100-fold with 0.05 mol/l Tris-HCl buffer (with or without 0.1% Triton X-100). Appropriate calibration curves were obtained in the ELISA by using apoB-48 recombinant antigen. RESULTS: No cross-reactivity (<0.001%) was found with apoB-100, as verified by ELISA and Western blot analyses. Intra- and inter-assay CVs were 4.8% and 5.4%, respectively. Recovery of added recombinant apoB-48 in serum was within 94-105%. The assay linearity was intact >5-fold dilution of serum by dilution buffer. ApoB-48 levels in healthy controls (n=18) at fasting were within the range of 2.69-6.56 microg/ml (mean+/-S.D.: 4.60+/-1.54 microg/ml). In healthy subjects, serum apoB-48 concentrations markedly increased in the postprandial state, in parallel with serum triglycerides. CONCLUSION: This method for measuring apoB-48 using the monoclonal antibody 4C8 is simple, reliable and suitable for routine analyses.
Authors: Jeffrey M Saland; Lisa M Satlin; Jeanna Zalsos-Johnson; Serge Cremers; Henry N Ginsberg Journal: Kidney Int Date: 2016-05-07 Impact factor: 10.612
Authors: André J Tremblay; Benoît Lamarche; Valéry Lemelin; Lizbeth Hoos; Suzanne Benjannet; Nabil G Seidah; Harry R Davis; Patrick Couture Journal: J Lipid Res Date: 2010-12-01 Impact factor: 5.922
Authors: Gissette Reyes-Soffer; Steve Holleran; Wahida Karmally; Colleen I Ngai; Niem-Tzu Chen; Margarita Torres; Rajasekhar Ramakrishnan; William S Blaner; Lars Berglund; Henry N Ginsberg; Catherine Tuck Journal: J Lipid Res Date: 2009-05-08 Impact factor: 5.922