31 research outputs found
Naturally occurring and therapy-induced antibodies to human granulocyte colony stimulating factor (G-CSF) in human serum.
No full textNaturally occurring and therapy-induced antibodies to human granulocyte colony stimulating factor (G-CSF) in human serum
DETECTION AND EPITOPE MAPPING OF IMMUNOREACTIVE HUMAN ENDOTHELIN-1 USING ELISA AND A SURFACE PLASMON RESONANCE-BASED BIOSENSOR
No full textA surface plasmon resonance-based biosensor (BIA technology) and enzyme- linked immunosorbent assays (ELISA) have been used for detecting and characterizing human endothelin (ET), a potent vasoactive 21 amino acid polypeptide. Antibodies produced against the isoform ET-1 and its C-terminal eptapeptide ET-115-21 have been characterized with respect to their binding capacity to the two isoforms ET-1 and ET-3, the non-secreted portion of the precursor molecule Big.ET-122-38, the C-terminal of ET-1, six analogues of ET-116-21 each containing a substitution with Ala of a single amino acid in positions 16-21, respectively, and three synthetic cyclic peptides mimicking the N-terminal pollion of ET-1. Antibodies reacting with ET-1 also bound to ET-116-21 and, with less affinity, to ET-3 but did not cross-react with Big. ET-122-38. Ala substitution in positions 16, 17 and 19 of ET-116-21 hardly affected the antibody binding capacity of ET-116-21, whereas Ala substitution of Asp18 Ile20 and, in particular, Trp21, inhibited its immunoreactivity. The C-terminus thus represents an immunodominant epitope in ET-1 and is important for antibody binding. Epitope mapping using as antibody pairs polyclonal anti-ET-1 and monoclonal anti-ET-115-21 antibodies indicated the presence of another immunogenic domain in the N-terminal portion of the molecule. There was excellent agreement between the epitopes determined using ELISA and BIA analyses.
A surface plasmon resonance-based biosensor (BIA technology) and enzyme-linked immunosorbent assays (ELISA) have been used for detecting and characterizing human endothelin (ET), a potent vasoactive 21 amino acid polypeptide. Antibodies produced against the isoform ET-1 and its C-terminal eptapeptide ET-115-21 have been characterized with respect to their binding capacity to the two isoforms ET-1 and ET-3, the non-secreted portion of the precursor molecule Big.ET-122-38, the C-terminal of ET-1, six analogues of ET-116-21 each containing a substitution with Ala of a single amino acid in positions 16-21, respectively, and three synthetic cyclic peptides mimicking the N-terminal portion of ET-1. Antibodies reacting with ET-1 also bound to ET-116-21 and, with less affinity, to ET-3 but did not cross-react with Big.ET-122-38. Ala substitution in positions 16, 17 and 19 of ET-116-21 hardly affected the antibody binding capacity of ET-116-21, whereas Ala substitution of Asp18, Ile20 and, in particular, Trp21, inhibited its immunoreactivity. The C-terminus thus represents an immunodominant epitope in ET-1 and is important for antibody binding. Epitope mapping using as antibody pairs polyclonal anti-ET-1 and monoclonal anti-ET-115-21 antibodies indicated the presence of another immunogenic domain in the N-terminal portion of the molecule. There was excellent agreement between the epitopes determined using ELISA and BIA analyses
Natural and therapy-induced anti-GM-CSF and anti-G-CSF antibodies in human serum.
No full textSerum samples were obtained from patients with lymphoid and plasma cell malignancies who received after chemotherapy human recombinant GM-CSF or G-CSF. Sera from some patients revealed the presence of anti-cytokine antibodies, particularly after repetitive cytokine injections. Antibody Fab binding in a saturable manner by ELISA and Western immune-blotting confirmed antibody specificity. Anti-cytokine antibodies were detected before the exogenous cytokine injections in some patients, but increasing antibody levels were found after one or subsequent treatments. Low levels of anti-GM-CSF and anti-G-CSF antibodies were also detected in a relatively large proportion (about 10-30%) of normal sera from different adult healthy volunteers who had never been treated before with exologous cytokines as well as from cord blood. EBV-immortalized cord blood derived B-cell cultures were also found to produce anti GM-CSF and/or anti-G-CSF antibodies with high frequency
NATURALLY OCCURRING AND THERAPY-INDUCED ANTIBODIES TO HUMAN GRANULOCYTE COLONY-STIMULATING FACTOR (G-CSF) IN HUMAN SERUM
No full textSera were obtained from two groups of patients. Group A included 7 patients
with low-grade non-Hodgkin's lymphoma treated with three or more cycles of
standard-dose chemotherapy and recombinant human granulocyte-colony stimu-
lating factor (rhG-CSF). The cytokine was administered to half the patients after
the ®rst chemotherapy cycle and to the other half after the second according to
a randomized design and then to all patients from the third chemotherapy cycle
on, until documented hemopoietic reconstitution. Group B included 3 patients
with high-grade non-Hodgkin's lymphoma, 1 patient with resistant Hodgkin's
disease, and 1 patient with multiple myeloma who received high-dose chemotherapy
and rhG-CSF. Anti-G-CSF antibodies were detected in the sera of 4
patients. Both immunoglobulin IgM and IgG antibodies were detected at low
levels in pretreatment sera from one group A patient. IgG antibody titers increased
markedly during the ®rst and second periods of G-CSF administration. IgG class
antibodies developed in 3 group B patients during the ®rst course of rhG-CSF administration.
Circulating anti-G-CSF antibodies did not seem to affect hematological recovery.
Low levels of anti-G-CSF antibodies were also detected in sera (15/135) from
different healthy adults and in sera (5/40) from umbilical cord blood. Saturable
antibody binding and competition enzyme-linked immunosorbent assay (ELISA) and
immunoblotting con®rmed antibody speci®city
Comparison of a conventional immunoassay (ELISA) with a surface plasmon resonace-based biosensor for IGF-1 detection in cows’ milk
No full textDetection and epitope mapping of immunoreactive human endothelin-1 using ELISA and a surface plasmon resonance based biosensor.
No full textStructure-activity analysis of C-terminal endothelin analogues
No full textSeveral synthetic endothelin (ET) analogues of the C-terminal ET hexapeptide (ET16-21) were analyzed by radio-receptor competition binding assays and biologic activity using both ETA and ETB receptor subtypes. In addition, we produced a hybridoma monoclonal antibody, anti-ET15-21, that appeared to crossreact with the entire ET molecule and was able to neutralize its biologic activity. Antibody binding was measured with competition enzyme-linked immunosorbent assays and a surface plasmon resonance-based biosensor (BIA technology). The ET16-21 moiety was modified with systematic replacement of each residue by alanine (Ala-scan). Whereas the C-terminal residues (Asp(18), Ile(20), and particularly Trp(21)) were very important for both receptor binding and immunologic activity, Ala substitution in positions 16, 17, and 19 hardly affected such activities. Analysis of another series of synthetic ET16-21 analogues with the His(16) residue replaced by a non-amino-acidic block confirmed that the last two C-terminal residues are essential for receptor and antibody binding, whereas the central region of this hexapeptide is much more tolerant to modification. However, a critical steric conformation of the active hexapeptide is necessary
Naturally occurring and therapy-induced antibodies to human granulocyte colony-stimulating factor (G-CSF) in human serum.
No full text
