]> BibSonomy publications for /author/Marcovina http://www.bibsonomy.org/burst/author/Marcovina BibSonomy BuRST Feed for /author/Marcovina 2008-07-20T23:48:18+02:00 Epitope mapping analysis of apolipoprotein B-100 using a surface plasmon resonance-based biosensor. http://www.bibsonomy.org/bibtex/264fee8a74054cacb021a4106ed8baf1f/biblio24 biblio24 2006-07-07T01:10:50+02:00 apob epitope mapping spr
Epitope mapping analysis of apolipoprotein B-100 using a surface plasmon resonance-based biosensor.
L. Laricchia Robbio and P. Uboldi and S. Marcovina and R. P. Revoltella and A. L. Catapano Biosens Bioelectron 16 963--969 (2001)
to apob epitope mapping spr by biblio24 on 2006-07-07 01:10:50 ]]> Institute of Mutagenesis and Differentiation, CNR, Area della Ricerca San Cataldo, 56100 Via Moruzzi, Ghezzano-Pisa, Italy. lel@imd.pi.cnr.itBiosens BioelectronDecember9-12963--969Epitope mapping analysis of apolipoprotein B-100 using a surface plasmon resonance-based biosensor.162001apob epitope mapping spr 2006-07-07 01:10:50.0Using a surface plasmon resonance (SPR)-based biosensor (BIA-technology), we have studied the interaction of ten different murine monoclonal antibodies (mAbs, all IgG(1)), raised against the main protein constituent of human low density lipoprotein (LDL), i.e. the apolipoprotein B-100 (apoB-100). These mAbs identify distinct domains on apoB-100, relevant to LDL-receptor interaction: epitopes in the amino-terminal region (mAbs L7, L9, L10 and L11: aa 1-1297) and in the middle region (mAb 6B: aa 1480-1693; mAbs 2A, 3B: aa 2152-2377; mAbs 9A, L2 and L4: aa 2657-3248) of native apoB-100. A multisite binding analysis was performed to further characterize the epitopes recognized by all these mAbs. A rabbit anti-mouse IgG(1)-Fc antibody (RAM.Fc) was first coupled to the gold surface in order to capture one anti-human apoB-100 mAb. ApoB-100 protein was subsequently injected and allowed to react with this immobilized, oriented antibody. Multisite binding assays were then performed, by sequentially flowing other mAbs, in different orders, over the sensing surface. The capacity of each mAb to interact with the entrapped apoB-100 in a multimolecular complex was monitored in real time by SPR. The results achieved were comparable to those obtained by western immunoblotting using the same reagents. However, SPR ensures a more detailed epitope identification, demonstrating that BIA-technology can be successfully used for mapping distinct epitopes on apoB-100 protein in solution dispensing with labels and secondary tracers; moreover, compared with conventional immunoassays, it is significantly time saving (CNR-P.F. MADESS 2). Monoclonal antibodies to human low density lipoprotein identify distinct areas on apolipoprotein B-100 relevant to the low density lipoprotein-receptor interaction. http://www.bibsonomy.org/bibtex/28470a55ffd3031830371a1e0072d15bf/biblio24 biblio24 2006-07-07T01:10:50+02:00 apob epitope mapping
Monoclonal antibodies to human low density lipoprotein identify distinct areas on apolipoprotein B-100 relevant to the low density lipoprotein-receptor interaction.
S. Fantappiè and A. Corsini and A. Sidoli and P. Uboldi and A. Granata and T. Zanelli and P. Rossi and S. Marcovina and R. Fumagalli and A. L. Catapano Journal of Lipid Research 33 1111--1121 (1992)
to apob epitope mapping by biblio24 on 2006-07-07 01:10:50 ]]> Institute of Pharmacological Sciences, University of Milano, Italy.Journal of Lipid ResearchAugust81111--1121Monoclonal antibodies to human low density lipoprotein identify distinct areas on apolipoprotein B-100 relevant to the low density lipoprotein-receptor interaction.331992apob epitope mapping 2006-07-07 01:10:50.0We have characterized the epitopes for ten murine monoclonal antibodies (Mabs) to human low density lipoprotein (LDL) and studied their ability to interfere with the LDL-receptor interaction. The epitopes for the antibodies were defined by using the following approaches: 1) interaction with apoB-48; 2) interaction with apoB-100 thrombolytic fragments; and 3) interaction with beta-galactosidase-apoB fusion proteins spanning different areas of the apoB-100 sequence. The results obtained are consistent with the following map of epitopes: Mab 6E, amino acids (aa) 1-1297, Mabs 5A and 6B, aa 1480-1693, Mabs 2A, 7A, 3B, and 4B, aa 2152-2377, Mabs 8A and 9A, aa 2657-3248 and 3H, aa 4082-4306. Four Mabs (2A, 5A, 7A, and 9A) whose epitopes are located in three different areas of apoB, dramatically reduced (up to 95%) the LDL-receptor interaction on cultured human fibroblasts; Fab fragments were as effective as the whole antibodies. Mab 3H, on the other hand, increased LDL binding up to threefold. These findings are consistent with the hypothesis that several areas of apoB-100 are involved independently or in concert in modulating the apoprotein B conformation required for interaction with the LDL receptor. Interaction of LDL, Lp[a], and reduced Lp[a] with monoclonal antibodies against apoB. http://www.bibsonomy.org/bibtex/2c2f02dd34a1c927e4d1496d639f195f1/biblio24 biblio24 2006-07-07T01:10:50+02:00 apob epitope mapping
Interaction of LDL, Lp[a], and reduced Lp[a] with monoclonal antibodies against apoB.
A. Gries and C. Fievet and S. Marcovina and J. Nimpf and H. Wurm and H. Mezdour and J. C. Fruchart and G. M. Kostner J Lipid Res 29 1--8 (1988)
to apob epitope mapping by biblio24 on 2006-07-07 01:10:50 ]]> Institute of Physiology, University of Graz, Austria.J Lipid ResJanuary11--8Interaction of LDL, Lp[a], and reduced Lp[a] with monoclonal antibodies against apoB.291988apob epitope mapping 2006-07-07 01:10:50.0Five monoclonal antibodies (2A, 9A, 6B, L3, L7) produced in mice against human apolipoprotein B were investigated by competitive and inhibitive electroimmunoassay (EIA) for their reactivity with low density lipoprotein (LDL), lipoprotein[a] (Lp[a]), and reduced Lp[a]. All of the antibodies reacted with apoB of the different lipoproteins indicated by very similar slopes of the binding curves. None of them gave a positive reaction with apolipoprotein[a]. The amount of apoB required for 50% inhibition of antibody binding varied for the different antibodies and lipoproteins. Antibody 9A showed almost the same affinity for LDL, Lp[a], and reduced Lp[a]. Antibodies 2A and 6B bound about twofold better to LDL and reduced Lp[a] than to untreated Lp[a]. Antibodies L3 and L7 needed nearly threefold higher amounts of Lp[a]-apoB for 50% inhibition of antibody binding than of apoB of LDL and reduced Lp[a]. The amount of apoB required for 50% inhibition of antibody binding was somewhat higher in inhibitive assay than in competitive assay. We suggest that apo[a] covers certain epitopes of apoB in native Lp[a] leading to a reduced reaction with the monoclonal antibodies. However, it could also be that the binding of the [a]antigen to apoB via disulfide bridges causes profound conformational changes of the apoB region exposed to the surface. Immunoreactivity of apo B towards monoclonal antibodies that inhibit the LDL-receptor interaction: effects of LDL oxidation. http://www.bibsonomy.org/bibtex/2acca9a5e11f5de28586667a664f1e859/biblio24 biblio24 2006-07-07T01:10:50+02:00 apob epitope mapping
Immunoreactivity of apo B towards monoclonal antibodies that inhibit the LDL-receptor interaction: effects of LDL oxidation.
S. Negri and P. Roma and R. Fogliatto and P. Uboldi and S. Marcovina and A. L. Catapano Atherosclerosis 101 37--41 (1993)
to apob epitope mapping by biblio24 on 2006-07-07 01:10:50 ]]> Institute of Pharmacological Sciences, University of Milano, Italy.AtherosclerosisJune137--41Immunoreactivity of apo B towards monoclonal antibodies that inhibit the LDL-receptor interaction: effects of LDL oxidation.1011993apob epitope mapping 2006-07-07 01:10:50.0We studied the immunochemical stability of the epitopes for six monoclonal antibodies to human apolipoprotein B-100 upon Cu(2+)-mediated (20 microM) oxidation of LDL. The antibodies used in this study, some of which are known to interfere with the interaction of LDL with their cellular receptors, recognize epitopes in the amino terminal region (Mb 19), in the middle part (6B, 2A, 7A, and 9A) and near aa 3500 (Mb 47) of native apo B. All antibodies except one (7A) recognized native and oxidized LDL (OxLDL) equally well; the immunoreactivity of the epitope for Ab 7A was markedly reduced upon LDL oxidation. Since antibodies 2A, 7A, 9A, and Mb 47 inhibit the LDL-receptor interaction and OxLDL poorly interact in vitro with the LDL receptor we conclude that: (1) various epitopes for monoclonal antibodies against native apo B are spared upon LDL oxidation; and (2) the epitopes for antibodies 2A, 9A, and Mb 47 do not define a unique domain of apo B directly involved in the binding of LDL to their receptor. Monoclonal antibody 5A binds apolipoprotein B-48 and inhibits the low density lipoprotein-receptor interaction. http://www.bibsonomy.org/bibtex/28a5794e6f329c353fcea2f5897cbd46b/biblio24 biblio24 2006-07-07T01:10:50+02:00 apob epitope mapping
Monoclonal antibody 5A binds apolipoprotein B-48 and inhibits the low density lipoprotein-receptor interaction.
A. Corsini and S. Fantappiè and S. Marcovina and A. Granata and R. Fumagalli and A. L. Catapano Biochem Biophys Res Commun 162 908--915 (1989)
to apob epitope mapping by biblio24 on 2006-07-07 01:10:50 ]]> Institute of Pharmacological Sciences, University of Milan, Italy.Biochem Biophys Res CommunAugust3908--915Monoclonal antibody 5A binds apolipoprotein B-48 and inhibits the low density lipoprotein-receptor interaction.1621989apob epitope mapping 2006-07-07 01:10:50.0In a panel of 10 monoclonal antibodies raised against human LDL we detected three antibodies (named 5A, 6B, and 6E) which recognize both apolipoprotein B-100 and B-48. Antibody 5A inhibited, in a dose dependent manner, the interaction of 125I-LDL with their receptor on human skin fibroblasts. Using thrombolytic fragments, the epitope of antibody 5A was mapped to the carboxy terminal region of apo B-48. MAB 5A was equipotent with MAB Mb 47, an inhibitory antibody whose epitope lies near a putative receptor binding domain of apo B in thrombolytic fragment T2. These findings suggest that areas other than the carboxy terminal portion of apo B-100 may participate in the LDL-receptor interaction, either directly or by determining the exposition of high affinity sites of apo B-100. Noncompetitive enzyme-linked immunoassay for apolipoprotein B in serum. http://www.bibsonomy.org/bibtex/24ccc8738651f6dc00121c376f01d16fa/biblio24 biblio24 2006-07-07T01:10:50+02:00 apob elisa
Noncompetitive enzyme-linked immunoassay for apolipoprotein B in serum.
C. Fievet and M. Koffigan and D. Ouvry and S. Marcovina and Y. Moschetto and J. C. Fruchart Clinical Chemistry 30 98--100 (1984)
to apob elisa by biblio24 on 2006-07-07 01:10:50 ]]> Clinical ChemistryJanuary198--100Noncompetitive enzyme-linked immunoassay for apolipoprotein B in serum.301984apob elisa 2006-07-07 01:10:50.0We used a noncompetitive enzyme-linked immunoassay to measure apolipoprotein B (apo-B) concentration in human plasma. Goat anti-lipoprotein B immunoglobulins were adsorbed to the surface of polystyrene balls. After washing, this solid-phase antibody was incubated with antigen (plasma from normal or hyperlipoproteinemic fasting subjects), washed, and then incubated with peroxidase-labeled goat anti-lipoprotein B IgG. After a last washing, we measured the bound label, which provided a direct measurement of the antigen. Under optimized assay conditions, the minimum detectable concentration was 50 ng per assay. The assay may be used to measure apo-B in different lipoprotein fractions (low- or very-low-density) and yields values that compared favorably with those obtained by electroimmunoassay (r = 0.86). The assay offers several advantages over existing techniques: sensitivity, specificity, simplicity, avoidance of radioisotopes, and potential for use with monoclonal antibodies. Monoclonal antibodies can precipitate low-density lipoprotein. I. Characterization and use in determining apolipoprotein B. http://www.bibsonomy.org/bibtex/2fce4002b3d7e009f7257217fc0d1656d/biblio24 biblio24 2006-07-07T01:10:50+02:00 antibody apob elisa
Monoclonal antibodies can precipitate low-density lipoprotein. I. Characterization and use in determining apolipoprotein B.
S. Marcovina and D. France and R. A. Phillips and S. J. Mao Clinical Chemistry 31 1654--1658 (1985)
to antibody apob elisa by biblio24 on 2006-07-07 01:10:50 ]]> Clinical ChemistryOctober101654--1658Monoclonal antibodies can precipitate low-density lipoprotein. I. Characterization and use in determining apolipoprotein B.311985antibody apob elisa 2006-07-07 01:10:50.0We produced 20 mouse monoclonal antibodies against human plasma low-density lipoprotein (LDL). Individually they failed to precipitate LDL in agarose gel by the double-immunodiffusion technique; collectively they did, or as few as two combined monoclonal antibodies could do so. To mimic polyclonal antibodies in determination of apolipoprotein B (apo B) by radial immunodiffusion, a combination of four particular monoclonal antibodies (clones A, B, C, and D) was necessary. We characterized these four clones with respect to temperature dependency, affinity, total binding to 125I-labeled LDL, and specificity to the different species of apolipoprotein B. Two monoclonal antibodies (B and C) bound 100% of 125I-labeled LDL; clones A and D bound 80% and 87%, respectively. All four clones bound maximally to LDL at 4 degrees C. The affinity constants for clones A, B, C, and D were 0.6, 2.1, 3.8, and 2.3 X 10(9) L/mol, respectively. By the Western blotting technique, the four monoclonal antibodies all reacted with the species B-100 and B-74 of apolipoprotein B, and to various degrees with B-48 and B-26. Radial immunodiffusion (chi) and direct enzyme-linked immunosorbent assay (y) with a mixture of the four monoclonal antibodies gave almost identical results for 70 patients: y = 0.921 chi-2.58; r = 0.933. Monoclonal antibodies can precipitate low-density lipoprotein. II. Radioimmunoassays with single and combined monoclonal antibodies for determining apolipoprotein B in serum of patients with coronary artery disease. http://www.bibsonomy.org/bibtex/2d4f7638f23ba1f990ce9519167a5ca8b/biblio24 biblio24 2006-07-07T01:10:50+02:00 antibody apob elisa epitope immunoassay
Monoclonal antibodies can precipitate low-density lipoprotein. II. Radioimmunoassays with single and combined monoclonal antibodies for determining apolipoprotein B in serum of patients with coronary artery disease.
S. Marcovina and B. A. Kottke and S. J. Mao Clinical Chemistry 31 1659--1663 (1985)
to antibody apob elisa epitope immunoassay by biblio24 on 2006-07-07 01:10:50 ]]> Clinical ChemistryOctober101659--1663Monoclonal antibodies can precipitate low-density lipoprotein. II. Radioimmunoassays with single and combined monoclonal antibodies for determining apolipoprotein B in serum of patients with coronary artery disease.311985antibody apob elisa epitope immunoassay 2006-07-07 01:10:50.0We have established four lines of monoclonal antibodies against human low-density lipoproteins (LDL) that, mixed in equal proportions, can precipitate LDL in gel and so can be used for apolipoprotein (apo) B determination in plasma. One monoclonal antibody (clone A), with a relatively low binding affinity to LDL (ka = 0.6 X 10(9) L/mol) and recognizing only two species of apo B, significantly underestimated the concentration of apo B in 74 patients with and 27 without coronary artery disease (CAD). High-affinity monoclonal antibody C (Ka = 3.8 X 10(9) L/mol), which recognized all four apo B species, gave the same value for apo B as determined with the mixture of monoclonal antibodies. The latter results (by radioimmunoassay, y) correlated well with those by radial immunodiffusion (chi): y = 0.994 chi + 0.003 (r = 0.987). The CAD patients showed an increased concentration of apo B as compared to the angiographically documented CAD-negative patients. Except for the values determined by clone B (p = 0.07), the increase was statistically significant (p = 0.002-0.018) for values determined by use of the other clones or their mixture. Development of a radial immunodiffusion technique employing monoclonal antibodies for apolipoprotein B determination in human plasma. http://www.bibsonomy.org/bibtex/2bba127a19be5bc9b2b2bc015a578c132/biblio24 biblio24 2006-07-07T01:10:50+02:00 antibody apob immunoassay
Development of a radial immunodiffusion technique employing monoclonal antibodies for apolipoprotein B determination in human plasma.
S. Marcovina and G. Di Cola and C. Rapetto and C. Fievet and V. Clavey and J. C. Fruchart Clinica Chimica Acta 147 117--125 (1985)
to antibody apob immunoassay by biblio24 on 2006-07-07 01:10:50 ]]> Clinica Chimica ActaApril2117--125Development of a radial immunodiffusion technique employing monoclonal antibodies for apolipoprotein B determination in human plasma.1471985antibody apob immunoassay 2006-07-07 01:10:50.0Twenty monoclonal antibodies obtained from two different fusions of SP2/0-Ag 14 cell line (non-secretor hybridoma) with human plasma low density lipoproteins have been selected. We found that a mixture formed by the 20 monoclonal antibodies was able to form a single precipitin line with human plasma low density lipoproteins by a double gel diffusion technique. Further studies revealed that only two monoclonal antibodies were needed to precipitate low density lipoproteins in gel. However, a minimum of four particular monoclonal antibodies was required to obtain an optimal precipitin ring and a linear standard curve within 24 h using a radial immunodiffusion technique. We have then compared the radial immunodiffusion performed with a monoclonal antibody mixture to those employing conventional goat and rabbit antibodies in terms of plasma apolipoprotein B determinations. The apolipoprotein B values determined by monoclonal antibodies significantly correlate with the values obtained by the technique using goat (r = 0.95; p less than 0.001) and rabbit (r = 0.95; p less than 0.001) antibodies. Our data indicate that a mixture of monoclonal antibodies can mimic conventional antibodies in terms of immunoprecipitation and apolipoprotein B determination. Effects of lyophilization of serum on the measurement of apolipoproteins A-I and B. http://www.bibsonomy.org/bibtex/22c5b4cd125fe6db9d53102db776d8543/biblio24 biblio24 2006-07-07T01:10:50+02:00 apob immunoassay methodology
Effects of lyophilization of serum on the measurement of apolipoproteins A-I and B.
S. M. Marcovina and J. L. Adolphson and M. Parlavecchia and J. J. Albers Clin Chem 36 366--369 (1990)
to apob immunoassay methodology by biblio24 on 2006-07-07 01:10:50 ]]> Northwest Lipid Research Center, Harborview Medical Center, Seattle, WA 98104-2499.Clin ChemFebruary2366--369Effects of lyophilization of serum on the measurement of apolipoproteins A-I and B.361990apob immunoassay methodology 2006-07-07 01:10:50.0A common accuracy-based standardization program is indispensable for establishing reference intervals for the clinical use of apolipoproteins. The development and distribution of reference materials and quality-control materials that do not exhibit matrix effects between methods is essential to the standardization process. We examined the suitability of lyophilized material as a common reference material for the measurement of apolipoproteins A-I and B. We determined values for apolipoproteins A-I and B in frozen and lyophilized serum pools, using different immunochemical approaches. We found little or no differences in apolipoprotein A-I values between frozen and lyophilized pools as determined by the different methods. In contrast, values for apolipoprotein B in lyophilized samples were consistently lower than those obtained for frozen samples. After adjusting for the effect of dilution due to reconstitution, the difference in the apolipoprotein B values for lyophilized as compared with frozen samples ranged from -26% to 4%, depending upon the assay method. Evidently, serum pools in lyophilized from are not a suitable matrix for reference materials for apolipoprotein B measurements but can be used for apolipoprotein A-I measurements.