Home LiteratureArticle Details
PMID: 308657 Published · ppublish English Journal Article

Isolation of myeloma variants with predefined variant surface immunoglobulin by cell sorting.

Liesegang B, Radbruch A, Rajewsky K

Abstract

We describe a procedure for the isolation of somatic cell variants in which gene products are expressed on the cell surface that are not expressed in the wild type. Cloned cells of the myeloma line MPC 11, which expresses an IgG2b protein, were incubated with an antiserum specific for IgGI and IgG2a. Cells reacting with this antiserum were stained with a fluorescent anti-antiserum and enriched in three cycles of sorting in the fluorescence-activated cell sorter and subsequent growth in vitro. From the enriched population two variants were isolated by cloning in soft agar. One of them expressed a variant immunoglobulin that types serologically as an IgG2a but whose variable portion was idiotypically related to that of the MPC 11 wild-type protein.

MeSH Terms
B-Lymphocytes/metabolism Cell Separation/methods Genetic Variation Immunoglobulin G/biosynthesis Immunoglobulin Idiotypes/genetics Multiple Myeloma/pathology Neoplasms, Experimental/pathology Receptors, Antigen, B-Cell/biosynthesis,genetics
Chemicals
Immunoglobulin G Immunoglobulin Idiotypes Receptors, Antigen, B-Cell
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Liesegang B
Radbruch A
Rajewsky K
References (16)
16 references, click to expand
  1. An investigation of closely related gamma-myeloma proteins and normal mouse gamma-globbulin by partial enzymic degradation and starch-gel electrophoresis.
    Nature. 1961 Jun 10;190:980-3 PMID: 13684802
  2. Somatic mutants and antibody diversity.
    Immunol Rev. 1977;36:51-72 PMID: 408265
  3. Idiotypic analysis of the response of C57BL/6 mice to the (4-hydroxy-3-nitrophenyl)acetyl group.
    Eur J Immunol. 1977 Aug;7(8):559-65 PMID: 409605
  4. Primary and secondary variants in immunoglobulin heavy chain production.
    Nature. 1976 Dec 2;264(5585):480-2 PMID: 826832
  5. The requirement of an adherent cell substratum for the growth of developing plasmacytoma cells in vivo.
    J Exp Med. 1976 Dec 1;144(6):1554-67 PMID: 1003103
  6. The inheritance of individual antigenic specificities of rabbit antibodies to streptococcal carbohydrates.
    J Exp Med. 1971 Aug 1;134(2):532-52 PMID: 4104426
  7. Isoelectric focusing in polyacrylamide gel and its application to immunoglobulins.
    Nature. 1968 Jul 6;219(5149):66-7 PMID: 4173351
  8. The cross-linking of proteins with glutaraldehyde and its use for the preparation of immunoadsorbents.
    Immunochemistry. 1969 Jan;6(1):53-66 PMID: 4179831
  9. A simplified procedure for the preparation of immunoglobulin-class-specific antisera.
    J Immunol. 1969 Jul;103(1):56-61 PMID: 4183706
  10. Analysis of cell populations with a fluorescence-activated cell sorter.
    Ann N Y Acad Sci. 1975 Jun 30;254:163-71 PMID: 52308
  11. The identification of mouse myeloma cells which have undergone mutations in immunoglobulin production.
    Soc Gen Physiol Ser. 1974;29:233-44 PMID: 4215140
  12. Fluorescence activated cell sorting.
    Rev Sci Instrum. 1972 Mar;43(3):404-9 PMID: 5013444
  13. The use of proteolytic enzymes for the mapping of structural rearrangements in the chromosomes of man.
    Chromosoma. 1972;36(2):204-10 PMID: 5016007
  14. Variants of a mouse myeloma cell line that synthesize immunoglobulin heavy chains having an altered serotype.
    Proc Natl Acad Sci U S A. 1975 Apr;72(4):1427-30 PMID: 805428
  15. Rate of somatic mutation in immunoglobulin production by mouse myeloma cells.
    Proc Natl Acad Sci U S A. 1971 Jan;68(1):219-23 PMID: 5276293
  16. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1978-08-00
Pages
3901-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC392896
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]