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Journal ArticleDOI

Continuous cultures of fused cells secreting antibody of predefined specificity

07 Aug 1975-Nature (Nature)-Vol. 256, Iss: 5517, pp 495-497

TL;DR: The derivation of a number of tissue culture cell lines which secrete anti-sheep red blood cell (SRBC) antibodies is described here, made by fusion of a mouse myeloma and mouse spleen cells from an immunised donor.

AbstractTHE manufacture of predefined specific antibodies by means of permanent tissue culture cell lines is of general interest. There are at present a considerable number of permanent cultures of myeloma cells1,2 and screening procedures have been used to reveal antibody activity in some of them. This, however, is not a satisfactory source of monoclonal antibodies of predefined specificity. We describe here the derivation of a number of tissue culture cell lines which secrete anti-sheep red blood cell (SRBC) antibodies. The cell lines are made by fusion of a mouse myeloma and mouse spleen cells from an immunised donor. To understand the expression and interactions of the Ig chains from the parental lines, fusion experiments between two known mouse myeloma lines were carried out.

Topics: Hybridoma technology (63%), Monoclonal antibody (57%), Cell culture (55%), Screening procedures (54%), Cell fusion (53%)

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Citations
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Journal ArticleDOI
TL;DR: This work speculates on the reasons behind this large discrepancy between the expectations arising from proteomics and the realities of clinical diagnostics and suggests approaches by which protein-disease associations may be more effectively translated into diagnostic tools in the future.
Abstract: The human plasma proteome holds the promise of a revolution in disease diagnosis and therapeutic monitoring provided that major challenges in proteomics and related disciplines can be addressed. Plasma is not only the primary clinical specimen but also represents the largest and deepest version of the human proteome present in any sample: in addition to the classical "plasma proteins," it contains all tissue proteins (as leakage markers) plus very numerous distinct immunoglobulin sequences, and it has an extraordinary dynamic range in that more than 10 orders of magnitude in concentration separate albumin and the rarest proteins now measured clinically. Although the restricted dynamic range of conventional proteomic technology (two-dimensional gels and mass spectrometry) has limited its contribution to the list of 289 proteins (tabulated here) that have been reported in plasma to date, very recent advances in multidimensional survey techniques promise at least double this number in the near future. Abundant scientific evidence, from proteomics and other disciplines, suggests that among these are proteins whose abundances and structures change in ways indicative of many, if not most, human diseases. Nevertheless, only a handful of proteins are currently used in routine clinical diagnosis, and the rate of introduction of new protein tests approved by the United States Food and Drug Administration (FDA) has paradoxically declined over the last decade to less than one new protein diagnostic marker per year. We speculate on the reasons behind this large discrepancy between the expectations arising from proteomics and the realities of clinical diagnostics and suggest approaches by which protein-disease associations may be more effectively translated into diagnostic tools in the future.

3,837 citations


Cites background from "Continuous cultures of fused cells ..."

  • ...The requirement for an isolated antigen was circumvented, however, following the introduction of monoclonal mouse antibodies by Kohler and Milstein (11) in 1975....

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Journal ArticleDOI
24 Mar 1988-Nature
TL;DR: A human IgGI antibody has been reshaped for serotherapy in humans by introducing the six hypervariable regions from the heavy- and light-chain variable domains of a rat antibody directed against human lymphocytes.
Abstract: A human IgGI antibody has been reshaped for serotherapy in humans by introducing the six hypervariable regions from the heavy- and light-chain variable domains of a rat antibody directed against human lymphocytes. The reshaped human antibody is as effective as the rat antibody in complement and is more effective in cell-mediated lysis of human lymphocytes.

3,133 citations


Journal ArticleDOI
TL;DR: It is proposed that one of the major long-term consequences of inadequate early nutrition is impaired development of the endocrine pancreas and a greatly increased susceptibility to the development of Type 2 diabetes.
Abstract: In this contribution we put forward a novel hypothesis concerning the aetiology of Type 2 (non-insulin dependent) diabetes mellitus. The concept underlying our hypothesis is that poor foetal and early post-natal nutrition imposes mechanisms of nutritional thrift upon the growing individual. We propose that one of the major long-term consequences of inadequate early nutrition is impaired development of the endocrine pancreas and a greatly increased susceptibility to the development of Type 2 diabetes. In the first section we outline our research which has led to this hypothesis. We will then review the relevant literature. Finally we show that the hypothesis suggests a reinterpretation of some findings and an explanation of others which are at present not easy to understand.

2,962 citations


Cites methods from "Continuous cultures of fused cells ..."

  • ...Further work, this time exploiting the monoclonal antibody technique [18], was required to devise assays with adequate specificity to resolve the complex mixture of insulin-like molecules present in plasma [19]....

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Patent
10 Jul 1991
Abstract: A member of a specific binding pair (sbp) is identified by expressing DNA encoding a genetically diverse population of such sbp members in recombinant host cells in which the sbp members are displayed in functional form at the surface of a secreted recombinant genetic display package (rgdp) containing DNA encoding the sbp member or a polypeptide component thereof, by virtue of the sbp member or a polypeptide component thereof being expressed as a fusion with a capsid component of the rgdp. The displayed sbps may be selected by affinity with a complementary sbp member, and the DNA recovered from selected rgdps for expression of the selected sbp members. Antibody sbp members may be thus obtained, with the different chains thereof expressed, one fused to the capsid component and the other in free form for association with the fusion partner polypeptide. A phagemid may be used as an expression vector, with said capsid fusion helping to package the phagemid DNA. Using this method libraries of DNA encoding respective chains of such multimeric sbp members may be combined, thereby obtaining a much greater genetic diversity in the sbp members than could easily be obtained by conventional methods.

2,740 citations


Journal ArticleDOI
29 Oct 1982-Science
TL;DR: Monoclonal antibodies specific for 5-bromodeoxyuridine have been produced and applied in detecting low levels of DNA replication on a cell-by-cell basis in vitro and do not cross-react with thymidine.
Abstract: Monoclonal antibodies specific for 5-bromodeoxyuridine have been produced and applied in detecting low levels of DNA replication on a cell-by-cell basis in vitro. The immunoglobulin-producing hybridomas were derived from spleen cells of mice immunized with a conjugate of iodouridine and ovalbumin. The cells were fused with the plasmacytoma line SP2/0Ag14. The antibodies produced are highly specific for bromodeoxyuridine and iododeoxyuridine and do not cross-react with thymidine. DNA synthesis in cultured cells exposed to bromodeoxyuridine for as short a time as 6 minutes can be detected easily and rapidly by an immunofluorescent staining method and quantitated by flow cytometry.

2,684 citations


References
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Journal ArticleDOI
26 Apr 1963-Science
TL;DR: Distinct plaques, each of which is due to the release of hemolysin by a single antibody-forming cell, are revealed by complement after incubation, in an agar layer, of a mixture of Sheep red cells and lymphoid cells from a rabbit immunized with sheep red cells.
Abstract: Distinct plaques, each of which is due to the release of hemolysin by a single antibody-forming cell, are revealed by complement after incubation, in an agar layer, of a mixture of sheep red cells and lymphoid cells from a rabbit immunized with sheep red cells.

2,270 citations


Journal ArticleDOI
14 Aug 1964-Science
TL;DR: Evidence can be obtained which suggests that mating may be followed by segregation in mice, and when two clonal lines of mouse fibroblasts are grown together for 4 days, hybrid cells can be detected by selective conditions.
Abstract: When two clonal lines of mouse fibroblasts, each containing a drug-resistant marker, are grown together for 4 days, hybrid cells can be detected by selective conditions. These hybrid cells are presumed to be the result of mating. By the same method evidence can be obtained which suggests that mating may be followed by segregation.

1,895 citations


Journal ArticleDOI
TL;DR: In this review of progress in immunology, an attempt is made to synthesize some of the rapidly accumulating observations in clinical medicine and experimental biology into a workable scheme, which may help pediatricians in their approach to the study and management of patients suspected of abnormalities in their resistance to infection.
Abstract: In this review of progress in immunology, an attempt is made to synthesize some of the rapidly accumulating observations in clinical medicine and experimental biology into a workable scheme, which may help pediatricians in their approach to the study and management of patients suspected of abnormalities in their resistance to infection. Like all hypothetical schemes, based on incomplete knowledge and imperfect interpretation, it must be considered tentative and useful mainly in establishing hypotheses to be demolished by future investigation. However, it may clarify the bewildering torrent of case reports, experimental observations and speculations which are pouring out in the current immunological and clinical literature. The roles of the phagocytic system, the thymus, the lymphocytes, and the plasma cells with their secretory products, the immunoglobulins, in defense against infection, as well as the clinical manifestations arising from deficiencies of each of these elements in the immunological system of the body, are discussed.

581 citations


Journal ArticleDOI
13 Feb 1965-Nature
TL;DR: Hybrid Cells Derived from Mouse and Man : Artificial Heterokaryons of Mammalian Cells from Different Species is presented, which describes how cells from different species can be hybrids.
Abstract: Hybrid Cells Derived from Mouse and Man : Artificial Heterokaryons of Mammalian Cells from Different Species

556 citations


Journal ArticleDOI
TL;DR: Assays of the growth inhibiting activities of thymidine and hydrocortisone show that the lymphoma cells are much more sensitive than the myeloma cells, suggesting that in these respects the cultured cell lines possess some of the differentiated characteristics of normal lymphocytes and plasma cells.
Abstract: The establishment and growth characteristics of nine different long-term continuous in vitro cell lines originating from five myeloma anad two lymphoma tumor lines of independent origin in BALB/c or C3H mice, is described. Different culture lines were initiated by somewhat different methods but they now all grow in fortified Eagle's medium supplemented with horse serum as stationary suspension cultures with doubling times between 16 and 26 h. Some of the lines are near tetraploid and contain marker chromosomes while others are diploid or near-diploid. Several of the myeloma lines synthesize and secrete immunoglobulin while others, probably as a consequence of mutation, do not. The two lymphoma lines may synthesize Ig but do not appear to secrete it. Assays of the growth inhibiting activities of thymidine and hydrocortisone show that the lymphoma cells are much more sensitive than the myeloma cells. There is evidence suggesting that in these respects, as well as others, the cultured cell lines possess some of the differentiated characteristics of normal lymphocytes and plasma cells. Possible uses of the cell lines to analyze at the biochemical level the modes of genetic control of such characteristics are discussed.

490 citations