Cloning and partial nucleotide sequence of human immunoglobulin mu chain cDNA from B cells and mouse-human hybridomas.

Dolby, Thomas W.; Devuono, J; Croce, Carlo M.

doi:10.1073/pnas.77.10.6027

Cited by 20 publications

(7 citation statements)

References 34 publications

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“…Although antibodies have been sequenced for almost four decades [ 3 ] and a very comprehensive protocol was released 20 years ago to isolate antibody sequences [ 4 ], a practical protocol for the design of CAR molecules does not seem to be available. We herein present a method that can be adapted in any lab to achieve the creation of a CAR.…”

Section: Introductionmentioning

confidence: 99%

Chimeric antigen receptor preparation from hybridoma to T-cell expression

Köksal

Baken

Warren

et al. 2019

Antibody Therapeutics

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The successful use of chimeric antigen receptor (CAR) for hematological cancer treatment has influenced the direction taken in translational research toward an increasing focus on personalized targeted immunotherapy. Thus, a growing number of labs worldwide are now interested in testing their old antibody collections in this format to broaden the spectrum of utility and improve safety and efficacy. We herein present a straightforward protocol for the identification of an antibody from a hybridoma and the design of the single chain fragment that will be placed on the extracellular part of the CAR construct. We further show how to test the expression and the activity of the construct in primary T cells. We illustrate our demonstration with two new CARs targeted against the B cell receptor, more precisely the light chains κ and λ, that represent potential alternatives to the CD19 CAR used in the treatment of B-cell malignancies. Statement of Significance Chimeric antigen receptor molecules are becoming increasingly popular, and an easy-to-follow method to isolate and express them is herein presented.

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Section: Introductionmentioning

confidence: 99%

Chimeric antigen receptor preparation from hybridoma to T-cell expression

Köksal

Baken

Warren

et al. 2019

Antibody Therapeutics

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“…Rabbitts and his coworkers have used this probe to demonstrate the synteny ofthe VH genes and the genes for heavy chains (8). The probe specific for the constant region of human 1 chain is a 655-base pair ,u cDNA that contains enough, sequences to code for amino acids glycine (residue 421) through the terminal tyrosine (residue 576) of the human ,u chain as well as the entire 3' noncoding sequence (27). The human y-specific probe was a kind gift ofJoel Bauxbaum (New York University).…”

Section: Methodsmentioning

confidence: 99%

Translocation of immunoglobulin VH genes in Burkitt lymphoma.

Erikson¹,

Nowell̀²,

Croce³

1982

Proc. Natl. Acad. Sci. U.S.A.

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178

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We have produced cell hybrids between mouse myeloma cells, which do not produce immunoglobulin chains, and Burkitt lymphoma cells (Daudi), which express surface IgM. Daudi Cells carry a reciprocal chromosome translocation between chromosomes 8 and 14, described as (8;14)(q24;q32). The hybrids were studied for the expression of human immunoglobulin chains and human isozyme markers, for the presence of human chromosomes, and for the presence of the human genes for heavy chain variable regions (VH) and mu and gamma chain constant (C) regions. The results indicate that the expressed mu chain gene is on normal chromosome 14 in Daudi cells. We have also determined that the chromosome 14 involved in the translocation (14q+) carries the gene for C mu and C gamma 1-4 and probably several genes for the variable region (V). Certain hybrids had lost both the chromosomes 14 but had retained the abnormal chromosome 8 (8q-) that carries the terminal end of the long arm of chromosome 14. These hybrids were studied for the presence of human VH, C mu,, and C gamma DNA sequences, and the results indicated that the hybrid cells with the 8q- chromosome contained VH genes that not C genes. Therefore, we conclude that, in the Daudi Burkitt lymphoma, the break in chromosome 14 occurred within the chromosome segment containing V region genes. As a result of the translocation some of these VH genes became associated with chromosome 8. It is possible that the expression of malignancy in Burkitt lymphoma is caused by immunoglobulin V region gene translocation resulting in activation of a gene on the long arm of human chromosome 8.

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“…On the contrary, lines CA46 and JD38 IV, which also carry a t(8;14) chromosome translocation, contain a c-myc gene rearranged with a C,, gene (14). In these two cell lines, the c-myc and C,, genes are located in the same 22-kb BamHI restriction fragment (33). Thus, breakpoint heterogeneity on both chromosomes 8 and 14 occurs in Burkitt lymphoma.…”

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confidence: 99%

“…The expression of human Ig chains was determined by immunoprecipitation of culture fluids or cytoplasmic extracts of the hybrid and parental cells using rabbit anti-human pu, K, A, 'y, and a chain-specific antibodies followed by the addition of 50 ,ud of a 10% suspension of fixed Staphylococcus aureus as described (11,12,26 (33), and a 1.2-kb EcoRI genomic probe that includes the first, the second, and part of the third exon ofthe human CM gene (unpublished data). The human y-specific probe was a kind gift of Joel Bauxbaum (New York University) (34).…”

mentioning

confidence: 99%