Application of chemically desialylated and degalactosylated human glycophorin for induction and characterization of anti-Tn monoclonal antibodies.

Human erythrocyte glycophorin was desialylated by mild acid hydrolysis and degalactosylated by Smith degradation. Two monoclonal antibodies (Tn5 and Tn56) obtained by immunization of mice with this 'artificial' Tn antigen were characterized and compared in some experiments with two antibodies (BRIC111 and LM225) obtained in other laboratories by immunization with Tn erythrocytes. The specific binding of the antibodies to glycophorins desialylated and degalactosylated on the nitrocellulose blot and to asialo-agalactoglycophorin-coated ELISA plates, and reactions with authentic Tn antigen served for identification of their anti-Tn specificity.

Analysis of peptidic epitopes recognized by the three monoclonal antibodies specific for the same region of glycophorin A but showing different properties.

Analysis of epitopes for the three monoclonal antibodies (GPA105, GPA33, OSK4-1) against glycophorin A (GPA) was performed with the use of proteolytic fragments of GPA, the synthetic nonapeptide with the sequence of amino acid residues 35-43 of GPA, and a series of peptides synthesized on plastic pins. The antibodies were specific for a short peptide sequence RAHE (a.a.

Monoclonal antibodies against human haptoglobin.

Three monoclonal antibodies: 2.36.71.

The Thomsen-Friedenreich antigen on human urothelial cell lines detectable by peanut lectin and monoclonal antibody raised against human glycophorin A.

The Thomsen-Friedenreich (TF) antigen is a cryptic disaccharide structure on human erythrocytes which can be exposed by neuraminidase treatment and which is supposed to be expressed in an unmasked form on some carcinoma cells. For its detection in addition to auto-, allo- and heteroantisera, PNA (peanut lectin) is being applied. In the present studies the mouse monoclonal antibody (MoAb) raised to asialoglycophorin from human erythrocytes was used.

Specific killing of mouse leukemic cells with ricin A-chain immunotoxin.

Monoclonal IgM antibody against L1210V leukemia was coupled with ricin A-chain using N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) as a cross--linking agent. The coniugate had potent concentration--dependent cytotoxicity against L1210V, L1210 and RL male 1 cells being completely non toxic to EL-4, P388, RPC-5 and mouse bone marrow cells. The minimum time required for killing L120V leukemia cells was 30h of in vitro exposure, at a concentration 10(-6) M (as assessed by trypan blue test).

Degradation of the human erythrocyte membrane band 3 studied with monoclonal antibody directed against an epitope on the cytoplasmic fragment of band 3.

The mouse hybridoma monoclonal antibody BIII.136 of the IgG2a class is specific for human erythrocyte band-3 protein. It was shown by means of immunoblotting and immunoprecipitation assays that the antibody recognized an epitope located in the cytoplasmic pole of the band-3 molecule within approximately 20 kDa from the N-terminal end.

Characterization of anti-prostatic acid phosphatase monoclonal antibody and its medical significance.

Monoclonal antibody to purified prostatic acid phosphatase from seminal plasma was produced by fusion of spleen cells from immunized mice with the Sp2/O-Ag 14 cell line. This hybridoma-derived antibody, designated MAb-14, was classified as IgG1 immunoglobulin. The apparent affinity constant of phosphatase.

Two monoclonal antibodies recognizing different epitopes of blood group N antigen.

Two monoclonal antibodies, N/61 (IgG2b) and N/92 (IgG2a), reacting preferentially with blood group N antigen, were obtained by immunization of BALB/c mice with human red cell membrane glycophorins. The antibodies showed distinctly different specificities. N/61 reacted with an epitope with NH2-terminal Leu residue, its free amino group, and sialic acid residues as essential components.

Immunoreactivity of proteolytic degradation products of human prostatic acid phosphatase.

Prostatic acid phosphatase (EC 3.1.3.

Genetic monitoring of inbred mouse strains maintained at the Bhabha Atomic Research Centre, Bombay: serological typing for H-2 haplotypes and lymphocyte differentiation markers.

Seven inbred mouse strains, AKR, CBA, C3H, C57BL/6, C57BR/cd, DBA/2 and Swiss, maintained at the Bhabha Atomic Research Centre, Bombay (designated Bh) were monitored for compatibility with standard strains as well as for genetical homogeneity. For this purpose, five individual mice from each strain were typed serologically for H-2 class I and class II antigens and for lymphocyte differentiation markers, Thy-2, Lyt-1, Lyt-2, TL and Ly-10. In this latter testing, 15 inbred strains from the Institute of Immunology and Experimental Therapy, Wrocław (designated Iiw) were included.

Detection of prostatic tissue and seminal plasma peptides reacting with human seminal fluid acid phosphatase antibodies.

IgG1 monoclonal antibody to purified seminal fluid phosphatase was raised by fusion of spleen cells from immunized mice with cell line Sp2/O-Ag 14 using simple method of screening for antiphosphatase antibody secreting clones. All molecular forms of catalytically active seminal fluid phosphatase and prostatic tissue phosphatase, resolved by chromatofocusing in pH gradient, react with this monoclonal antibody and with rabbit antiserum to purified seminal fluid phosphatase. Peptides of Mr 25,000 to 76,000 and of Mr 13,000 to 76,000 were adsorbed from the prostatic tissue extract and from seminal plasma on the monoclonal antibody-Sepharose column.

Quantitative analysis of tissue distribution of Ly10-like murine alloantigen(s) with antisera and monoclonal antibodies.

Tissue distribution of non-Lyt1.1 ("Ly10-like") antigen or antigens encoded by short chromosomal segment differentiating B6-Ly-1a congenic strain from B6 strain of mice was studied by quantitative absorption of (BALB/c X B6)F1 anti B6-Ly-1a antiserum and by direct cytotoxicity of Ly-10-132-12-26 monoclonal antibody on lymphoid cell populations. Identical strain but not tissue distribution pattern does not allow to conclude whether antiserum and monoclonal antibody detect the same or closely linked antigens.

Cell surface antigens of L-1210 leukemia recognized by monoclonal antibodies.

Conventional antisera to L-1210 leukemia were being prepared in our laboratory for nearly a decade and consistently the only specificity detectable was anti Mammary Leukemia antigen (ML). Serological analysis of five monoclonal antibodies obtained following the same immunization schedule showed more diverse pattern of reactivity. Two antigens detected belong to oncofetal category.

Monoclonal anti-TL antibody recognizing a TL.2-like antigen.

We obtained a monoclonal anti-TL antibody-producing hybridoma by fusion of Sp2/0 hybrid cells with spleen cells from (B6 X A-Tlab) F1 mice immunized with ASL1 leukemia cells. The antibody, TL-22-27-17, reacts in complement-dependent cytotoxicity assay with ASL1 cells as well as with normal thymocytes of TL+ mouse strains (B6-Tlaa, A, BALB/c, DBA/2 and 129) but not with thymocytes from TL- mice (B6--Ly-1a). In addition, it did not react with lymph node cells or Concanavalin A-induced spleen blasts from B6-Tlaa mice; this result shows that TL-22-27-17 recognizes TL and not Qa-1 antigen.

Serological identification of viral and virus-related antigens on DBA/2 mouse leukemia lymphocytes.

Allogenic, semisyngeneic and syngeneic sera of animals immunized with ML-positive leukemia L1210 cells, besides anti-ML antibodies, contain antibodies which react with Gross cellular surface antigen. ML antigen and Gross cellular surface antigen were shown by the immunoferritin test in electron microscopy, and by the blocking test, to be situated on different parts of the cell surface. No budding viral particles were found on the areas occupied by these antigens.