PubMed Health⌕ Search

Biomedical subjects

T Michaelsen

Publications and source records attributed to T Michaelsen.

4 recordsLinked to original sources
PubMed1992

B-cell hybridoma as intraperitoneal tumor model: correlation between tumor growth and monoclonal antibody production.

Murine B-cell hybridoma cells producing an immunoglobulin G1 (K13), specific for human immunoglobulin kappa chains were inoculated intraperitoneally in mice. After intraperitoneal injection of 10(6) K13 hybridoma cells, superficial intraperitoneal implants and ascites developed, resulting in death after 10 +/- 3 days (mean +/- SD). An immunoradiometric assay was developed to measure K13 in murine blood, ascites and culture supernatant. The assay utilized polymer beads coated with human immunoglobulin G. The amount K13 bound to the particles was measured with a 125I-labelled monoclonal rat antibody (LO-MG1-13) specific for mouse IgG1. The assay could be used over a wide working range (2-500 micrograms/l). Kinetic studies suggested that about 10(5) secreting cells were required for detection of K13 in blood. After injection of 10(6) cells, K13 was measurable in blood 1 day later in all animals. Nine of 33 mice injected with 10(5) or less cells survived, and initially showed rising K13 blood levels followed by decreasing blood levels. In conclusion, a close relationship was established between i.p. growth of the hybridoma K13 cell line and the MAb blood levels. The basic concepts of this assay can readily be adopted for other clones with the limitation that pure antigen is needed for solid phase extraction of the MAb from mouse blood.

Animals↗
PubMed1991

Shared idiotypic determinant in mono- and polyclonal anti-phospholipid antibodies with lupus anticoagulant activity.

Of 42 purified human myeloma proteins tested, two (IgG3 Her and IgM Mag) were found to possess strong lupus anticoagulant (LA) and anti-cephalin activity, as assessed by a dilute activated partial thromboplastin time (dAPTT) and ELISA test, respectively. For these proteins, we confirmed the observation reported by others that LA activity is present in the antigen-binding (Fab) portion of the immunoglobulin molecule. Rabbit anti-idiotype antibodies against IgG3 Her inhibited the anti-cephalin activity of this protein, suggesting that the anti-cephalin activity of IgG3 Her depends on the hypervariable part of the immunoglobulin and thus most probably is a true antigen-antibody reaction. The anti-Her idiotype antibodies were also able to bind to and inhibit the anti-cephalin activity of IgM Mag. ELISA binding and inhibition experiments showed that the anti-idiotype antiserum contained at least two sets of anti-idiotypes; one set that recognizes a cross-reactive idiotype shared by IgG3 Her and IgM Mag, and another set that seems to be unique to the immunizing protein IgG3 Her. Both sets of anti-idiotype antibodies also bound weakly to polyclonal (patient) IgG, indicating an idiotypic cross reaction.

Antibodies, Anti-Idiotypic↗
PubMed1977

The interaction of immunoglobulin heavy and light chains in the absence of the VH domain.

Recent studies of the micron- and kappa-chains of the first patient (GLI) with micronHCD indicated that the observed defect was the result of the failure of assembly of the intact kappa-chain to the micron-chain, which lacked the VH domain but had the CH1 Cys normally linked to the light chain. To explore the possibility that the VH region is necessary for the formation of the HL disulfide bond, in vitro studies were performed with GLI micron- and kappa-chains and with the CH1 domain and kappa-chain derived from an IgG3 myeloma protein, KUP, which yields separate VH, CH1, and kappa-chains after papain digestion and reduction. The proteins were reduced and allowed to reoxidize, and the combination products were assessed by gel chromatography under dissociating conditions by SDS-PAGE and by immunoprecipitation techniques. The results suggest that, although in vitro covalent and noncovalent combinations are possible between intact light chains and their autologous heavy chains even in the absence of the VH domain, the efficiency is less than that when the intact Fd region is used. Hence, it seems likely that lack of VH alone is not sufficient to explain the failure of assembly observed in muHCD.

Binding Sites, Antibody↗