ISSN:0272-457X    

DOI:10.1089/hyb.1983.2.1

年代:1983

数据来源: MAL

摘要:

Of several human fusion partners available for the production of monoclonal antibodies, only SKO-007 and RPMI 8226 have phenotypic features characteristic for myeloma cells. Cells from both lines exhibited abundant rough endoplasmic reticulum (RER) with a prominent Golgi apparatus, few free ribosomes, condensed nuclear chromatin, and absence of the Epstein-Barr virus determined nuclear antigen (EBNA). However, following prolonged passage of these lines, the amount of immunoglobulin (Ig) production has declined. The other cell lines, GM 1500 6TG-A11, KR-4, B6, HS-Sultan, and Raji possessed the phenotypic characteristics of B-lymphoblastoid cell lines (LCLs) and B-lymphomas including surface Ig expression, sparse RER, free polyribosomes, a poorly developed Golgi apparatus and strong EBNA expression. Accordingly, they secreted little (nanograms) or no Ig. However, hybrids constructed with two LCLs secrete very large amounts of Ig despite their expressed morphologic similarity to the parental lines. These data indicate that morphology can still be used as an important consideration in choosing a human fusion partner but other parameters such as fusion frequency, cloning efficiencies, and growth rates may be equally important.

ISSN:0272-457X    

DOI:10.1089/hyb.1983.2.7

年代:1983

数据来源: MAL

摘要:

Two human B lymphocyte-associated antigens, B1 and B2, were studied by immunoprecipitation using monoclonal antibodies. B1 was found to be a nonglycosylated protein of MW 35 kD, phosphorylated at serine and threonine. B2, a 140 kD MW antigen, was characterized as a glycoprotein without phosphorylated sites. Both proteins were found to contain portions which could be labeled with125I-iodonaphthylazide. The B1 protein displayed hydrophobic properties whereas B2 had a more amphiphilic character.

ISSN:0272-457X    

DOI:10.1089/hyb.1983.2.17

年代:1983

数据来源: MAL

摘要:

Spleen cells from a DBA/2 mouse immunized with RL♂1 tumor cells, a radiation induced BALB/c T-cell leukemia, were hybridized with the nonsecretor myeloma line NS.1. Four established hybrid cell lines continuously secreted antibodies that recognized a new alloantigenic specificity, tentatively called Ly-m22. This antigen is detectable on nearly 60% of lymph node cells, and 30% of spleen cells by direct cytotoxicity assay, but did not lyse significant number of cells of thymus and bone marrow. By absorption test, these lymphoid organs, i.e., lymph node, spleen, thymus, and bone marrow, were shown to express Ly-m22 determinant. The newly found antigen is expressed predominantly on T-cells. Analysis of BXD and SWXL recombinant inbred strains revealed close linkage betweenLy-m22andLtw-4loci on chromosome 1. The estimated recombination frequency is 0.027 ± 0.0

ISSN:0272-457X    

DOI:10.1089/hyb.1983.2.29

年代:1983

数据来源: MAL

摘要:

BALB/c mouse splenocytes from mice immunized with cells of the human hepatoma line Hep G2 were fused with SP2/0-Ag 14 mouse myeloma cells. Two monoclonal antibodies recognizing antigenic determinants (Hag-1, Hag-2) of hepatoma cell surface molecules were investigated. Analysis of immunoprecipitates by sodium dodecyl sulfate (SDS) gel electrophoresis revealed that the Hag-1 antigenic determinant is born on a 115, kD MW glycoprotein, and that the second antibody immunoprecipitates a group of surface proteins with MW of 230 kD, 79 kD, 23 kD, and 20 kD from human hepatoma cells. These antigenic determinants are present on cell lines derived from other human tumors, thus neither of the antibodies is hepatoma-specific; cross-reactivity with human colorectal carcinoma and some mammary carcinoma cell lines is notable. Using indirect immunofluorescence on frozen sections Hag-1 was detected in one of three liver biopsies tested whereas Hag-2 was demonstrated in all three. Both antigens were detected in sections of human kidney with Hag-2 localized to the proximal tubules.

ISSN:0272-457X    

DOI:10.1089/hyb.1983.2.39

年代:1983

数据来源: MAL

摘要:

We have obtained cell lines producing monoclonal antibodies against μ, α, λ, and λ1by fusing the mouse myeloma P3X63Ag8.653 with spleen cells from Lewis rats immunized with several λ1bearing myeloma proteins. The anti-λ1antibody is particularly useful. It was shown to be suitable for radioimmunoassays, for ELISAs when coupled to alkaline phosphatase, and for indirect fluorescence staining of the membrane as well as the cyto

ISSN:0272-457X    

DOI:10.1089/hyb.1983.2.49

年代:1983

数据来源: MAL

摘要:

Three rat anti-mouse monoclonal antibodies reactive with different populations of cells in mouse bone marrow have been extensively characterized with respect to their binding to cells in the lymphohemopoietic system. One antibody identifies lymphocytes committed exclusively to the B lineage. A second antibody identifies cells of the myeloid lineage including granulocytes and macrophages. The third antibody binds to nearly all of the bone marrow cells including erythrocytes, however, quantitative differences in antigenic expression allows the discrimination between early and late erythroid cells and can be used to purify eosinophils. The differential binding of these three monoclonal antibodies to mature cells, immature cells and progenitor cells (CFU-S, CFU-C (GM), BFU-E, and CFU-E) permits the construction of a scheme of lymphohemopoietic differentiation based on the expression of three cell surface antigens.

ISSN:0272-457X    

DOI:10.1089/hyb.1983.2.55

年代:1983

数据来源: MAL

摘要:

Monoclonal antibody A2B5 reacts with the cell surface of a series of amine precursor uptake decarboxylation (APUD)(1)cells and their tumors in many vertebrate species including chicken, rat, mouse, and man. We have studied the in vivo and in vitro binding of iodinated monoclonal antibody A2B5 to rat insulinoma cells. In vitro, radiolabeled A2B5 binds specifically to RINmSF insulinoma cells and the binding of125I-A2B5 is inhibited by unlabeled A2B5 or a ganglioside extract of RINm5F cells. In vivo, scintigrams taken Day 0 to Day 5 after injection of131I-labeled A2B5 showed a striking localization of131I-A2B5 in transplanted RIN tumors grown in syngeneic rats. Other control radiolabeled monoclonal antibodies did not concentrate in the tumors. 131I-labeled A2B5 did not concentrate in other transplantable tumors (colon adenocarcinoma, osteosarcoma, renal cell carcinoma, and bladder transitional cell carcinoma) grown in nude mice. The tumor/blood ratio detected 5 days after antibody injection, was approximately two to 12 times higher in the insulinoma compared to other organs and only in the insulinoma did131I-A2B5 show a higher concentration than control antibody125I-P3X63.

ISSN:0272-457X    

DOI:10.1089/hyb.1983.2.69

年代:1983

数据来源: MAL

摘要:

Spleen cells from BALB/c mice, immunized with SDS-acrylamide gel purified human fibroblast interferon (HuIFN-β) were fused with mouse myeloma cells. Culture fluids from the resulting hybrid cells were screened for HuIFN-β specificity by the following tests: (1) a solid-phase RIA using partially purified HuIFN-β, (2) a protein-transfer RIA using electrophoretically resolved and immobilized HuIFN-β, and (3) a bioassay which tests residual HuIFN-β activity in the supernatant following immunoprecipitation. Six HuIFN-β-specific hybridomas were identified; two were capable of partially neutralizing HuIFN-β activity. All six antibodies bind to the β1-IFN polypeptide synthesized inE. colicells containing a cloned β1-IFN DNA sequence. All six monoclonal antibodies were found to b

ISSN:0272-457X    

DOI:10.1089/hyb.1983.2.79

年代:1983

数据来源: MAL

摘要:

Data presented indicate that polyethylene glycol 1000 to be used for hybridoma production should be odorless at the time of purchase and should be stored at reduced temperatures in the absence of light. The optimal method of sterilization is by pressure filtration of the diluted PEG solution.

ISSN:0272-457X    

DOI:10.1089/hyb.1983.2.87

年代:1983

数据来源: MAL