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K Erb

Publications and source records attributed to K Erb.

69 records · Page 4Linked to original sources
PubMed1986

Measurement of C3 conversion by ELISA estimation of neo-determinants on the C3d moiety.

An ELISA assay estimating neo-determinants on the C3d moiety is described. The C3d neo-determinants were detected by incubating the sample on F(ab')2 anti-C3d-coated plates followed by development with biotin-labelled anti-C3d and enzyme-labelled avidin. The assay was compared with rocket immunoelectrophoresis (IE) and crossed IE. Serum from a factor I deficient patient showed no detectable C3d when analysed by rocket IE, but activation was evident when analysing by crossed IE or ELISA. The results suggest that the neo-determinants detected by ELISA become exposed when C3 is split into C3a and C3b and this interpretation was supported by kinetic analysis of C3 activation by the three methods.

Complement Activation↗
PubMed1986

Human-human hybridomas for the study of anti-tumor immune response in patients with colorectal cancer.

Human-human hybridoma technology was evaluated for the study of humoral immune reactions of colorectal cancer patients against their own tumors. Six fusions were carried out with lymphocytes from mesenteric lymph nodes from patients with colorectal cancer, using the human B-lymphoma cell line LICR-LON-HMy-2 as fusion partner. A total of 294 wells with cell growth were obtained. Supernatants from 26 of these reacted in enzyme-linked immuno-sorbent assay (ELISA) with one or more colon cancer cell lines. Cells from only one of these wells (D 4213) could be cloned. The clone was shown to produce antibody which by immunocytochemical analysis reacted with a panel of colon cancer cell lines and melanoma cell lines but not with several other cancer cell lines or normal human leukocytes. By immunohistochemical analysis on formalin-fixed paraffin-embedded tissue this antibody reacted strongly with antigen expressed by autologous and allogeneic colorectal cancers. Faint staining could occasionally be observed on normal colon epithelium. D4213 is a hybrid cell line since it is tetraploid and produces kappa and lambda light chains as well as gamma and mu chains, whereas HMy-2 produces only kappa and gamma chains. The study produces only kappa and gamma chains. The study suggests that patients may possess B cells producing antibody reactive with their own malignant cells.

Adenocarcinoma↗
PubMed1985

Increased non-specific binding of heat-treated proteins to plastic surfaces analyzed by ELISA and HPLC-fractionation.

Heat-treatment of proteins from human sera and bovine serum albumin caused an increase in their binding to microplates in a temperature-dependent manner. Quantitative data were obtained for 125I-labelled human IgG, with binding of up to 400 ng of 56 degrees-treated IgG per untreated well. However, only a minor increase in available antibody activity of adsorbed rabbit antibody was found upon pretreatment at 56 degrees. The increased binding to microplates as a result of pre-incubation at raised temperatures was by high pressure liquid gel permeation chromatography demonstrated to be accompanied by polymerization. Inhibition of the direct binding of the proteins to plastic surface was achieved most efficiently by coating with non-interfering proteins followed by non-ionic detergent (Tween 20), which should also be present in the wash buffer.

Animals↗
PubMed1984

Semi-automatic analysis of proteins and protein complexes by automated enzyme immunoassay after separation by high-performance gel-permeation chromatography. Size distribution of C3-IgG complexes.

Enzyme immunoassay (ELISA) was applied in the establishment of an immunological method for the characterization of column effluents. Serum samples, containing aggregated immunoglobulin (IgG), were separated by high-performance gel-permeation chromatography on a TSK column and fractions were collected directly in 96-well ELISA microplates. IgG was determined on anti-IgG-coated plates, followed by development with biotin-labelled anti-IgG and enzyme-labelled avidin. Complement factor C3 was determined on anti-C3-coated plates, developed with biotin-labelled anti-C3 and enzyme-labelled avidin. Complexes between IgG and complement factor C3 were determined on anti-IgG-coated plates, developed with biotin labelled anti-C3 antibody and enzyme-labelled avidin. Complex formation between C3 and IgG after incubation of serum with aggregated IgG was demonstrated. The methodology described is generally applicable to the analysis of biological components and is uniquely useful for the analysis of complexes between different components.

Animals↗
PubMed1984

Immune complex formation analysed by high-performance size exclusion chromatography (HPLC-SEC) using either 125I-labelled antigen or enzyme-linked immunosorbent assay (ELISA) for detection.

A physical/immunochemical method has been developed for the analysis of soluble immune complexes. Human serum with high amount of antibody against bovine serum albumin (BSA) was incubated at 37 degrees with a wide range of 125I-labelled BSA and separated according to size by high-performance size exclusion chromatography (HPLC-SEC) on a TSK G 6000 PW column. Fractions were collected onto microplates and analysed either by gamma counting or by an enzyme-linked immunosorbent assay (ELISA) technique detecting anti-BSA antibody. The size of the immune complexes were the same when analysed by the two methods. The heaviest immune complexes were seen in moderate antibody excess where the majority of the complexes were eluted in a broad peak around Mr of 7 X 10(6). In large antibody excess the immune complexes eluted at around 3 X 10(6). Around equilibrium two peaks were seen corresponding to 3 X 10(6) and 5 X 10(5). In antigen excess the peak at 3 X 10(6) is diminished and free antigen appears. No changes in the distribution of the complexes were observed when the serum was made 20 mM in EDTA before incubation with BSA.

Adult↗
PubMedDate not supplied

The role of Gilbert's syndrome and frequent NAT2 slow acetylation polymorphisms in the pharmacokinetics of retigabine.

Retigabine (RGB) is an investigational antiepileptic drug, which undergoes extensive UGT1A1, 1A9 and 1A4-mediated N-glucuronidation and N-acetylation. The mono-acetylated metabolite of RGB has some pharmacological activity and is denoted AWD21-360. We investigated whether the pharmacokinetics (PK) of RGB and AWD21-360 are altered in subjects with Gilbert's syndrome (GS) and/or with frequent N-acetyltransferase 2 (NAT2) slow acetylator (SA) polymorphisms. Based on consistent genotyping and phenotyping screening results, 37 Caucasian subjects (21-46 years; 31 men, six women) were assigned to one of the following groups: (1) absence of GS (non-GS)/rapid acetylator (RA) (N=11); (2) GS/RA (N=8); (3) non-GS/SA (N=11); (4) GS/SA (N=7). Subjects received single and multiple (b.i.d.) 200-mg oral RGB doses over 5 days. Blood samples were collected up to 60 h after dosing for plasma PK of RGB and AWD21-360. Group comparisons were performed by ANOVA. Single-dose PK of RGB and AWD21-360 and multiple-dose PK of RGB did not differ significantly between groups. After multiple dose treatment, RA subjects showed a significantly higher total exposure to AWD21-360 of about 32% (95% CI 101.9-172.5) relative to SA subjects (P=0.0362). The UGT1A1 metabolic capacity (i.e. presence or absence of GS), however, did not significantly affect the overall exposure to AWD21-360. The results indicate that the PK of RGB is unaltered in individuals with GS, in subjects with NAT2 SA status, and in carriers of both variants, whereas the total exposure to AWD21-360 is significantly related to the RA or SA status of subjects. Results further suggest that metabolic switching to the mono-acetylated metabolite AWD21-360 may partially compensate for the impaired glucuronidation capacity in GS subjects. RGB treatment showed no significant differences in tolerability and safety between groups.

Acetylation↗