Chimeric B72.3 mouse/human (IgG1) antibody directs the lysis of tumor cells by lymphokine-activated killer cells

Primus, F. James; Pendurthi, T. K.; Hutzell, Paula; Kashmiri, S. V. S.; Slavin, D.; Callahan, R J; Schlom, Jeffrey

doi:10.1007/bf01741406

Cited by 10 publications

(7 citation statements)

References 37 publications

(56 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Chimeric light and heavy chain cDNAs were constructed by fusing the variable regions of mT84.66 in-frame with the human constant regions from cB72.3 (34). For plant expression, the light chain DNA fragment was placed downstream of the ⍀ 5Ј UTR and the sequence encoding the codon optimized leader peptide LPL (Fig.…”

Section: Resultsmentioning

confidence: 99%

Transient expression of a tumor-specific single-chain fragment and a chimeric antibody in tobacco leaves

Vaquero

Sack

Chandler

et al. 1999

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

221

139

View full text Add to dashboard Cite

To evaluate the expression of different forms of a tumor-specific antibody in plants, we adapted a recently described Agrobacterium-mediated transient expression system. A recombinant single-chain Fv antibody (scFvT84.66) and a full-size mouse͞human chimeric antibody (cT84.66) derived from the parental murine mAb T84.66 specific for the human carcinoembryonic antigen were engineered into a plant expression vector. Chimeric T84.66 heavy and light chain genes were constructed by exchanging the mouse light and heavy chain constant domain sequences with their human counterparts and cloned into two independent plant expression vectors. In vivo assembly of full-size cT84.66 was achieved by simultaneous expression of the light and heavy chains after vacuum infiltration of tobacco leaves with two populations of recombinant Agrobacterium. Upscaling the transient system permitted purification of functional recombinant antibodies from tobacco leaf extracts within a week. His6-tagged scFvT84.66 was purified by immobilized metal affinity chromatography and cT84.66 by protein A affinity chromatography. Sufficient amounts of recombinant antibodies were recovered for detailed characterization by SDS͞PAGE, Western blotting, and ELISA.Monoclonal antibodies are essential tools in biology, biochemistry, and medicine. Their high affinity and specificity make them invaluable for diagnostic and therapeutic applications. However, the therapeutic use of murine mAbs is limited because they elicit a human anti-mouse antibody response and large amounts of antibody are required for therapy. These limitations may be overcome by engineering humanized antibodies and by producing these proteins in plants.The human anti-mouse antibody response can be reduced by using recombinant antibody (rAb) technology to replace the murine light and heavy chain constant domains with the corresponding human domains and the remaining murine variable domains to maintain the antigen specificity and affinity of the original mAb. A second approach is to use single-chain Fv antibody fragments (scFvs) where the constant domains have been removed and the variable domains are joined by a flexible linker (1). Compared with the full-size antibodies, scFvs display better tumor penetration and faster serum clearance but exhibit no effector functions. A critical step in testing the potential therapeutic use of these molecules is the development of a reproducible and efficient method for large-scale antibody production.Plants are potentially the most economical system for largescale production of rAbs (2, 3). rAbs are efficiently folded and assembled within the endoplasmic reticulum (ER) of plant cells (4-6) and retain the antigen binding properties of the antibodies produced by plasma or hybridoma cells (2,5,(7)(8)(9). Since the first report of antibody expression in transgenic plants (7), different engineered antibodies have been produced successfully, including full-size antibodies (8-11), Fab fragments (12), scFvs (13-21), and single-domain antibodies (22).Regene...

show abstract

Section: Resultsmentioning

confidence: 99%

Transient expression of a tumor-specific single-chain fragment and a chimeric antibody in tobacco leaves

Vaquero

Sack

Chandler

et al. 1999

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

221

139

View full text Add to dashboard Cite

show abstract

“…Fc-linked glycosyl residues are also implicated in complement fixation (8) MATERIALS AND METHODS Cell Lines and mAbs. The LS-174T human adenocarcinoma cell line, the A375 human melanoma cell line, the SP2/0-Agl4 murine myeloma cell line, and the KLE-B human endometrial carcinoma cells, which constitutively express TAG-72 antigen, have been described (10,13). The generation and properties ofmurine mAb CC49 (yl) have been described (1).…”

mentioning

confidence: 99%

“…cmAb CC49 (yl) was purified from bioreactor tissue culture fluid supplied by John Coleman (Dow Chemical, Midland, MI). The sources of MOPC-21 murine myeloma IgGl protein and purified human polyclonal IgG have been described (13).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Secretion of a single-gene-encoded immunoglobulin from myeloma cells.

Shu

Schlom

et al. 1993

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

View full text Add to dashboard Cite

We describe construction of a single gene encoding a single-chain immunoglobulin-like molecule. This single-gene approach circumvents inefficiencies inherent in delivering two genes into a mammalian cell and in the assembly of a functional immunoglobulin molecule. It would also facilitate ex vivo transfection of cells for gene-therapy protocols. SP2/0 murine myeloma cells transfected with the single gene SGACLCH1 expressed a single-chain protein, SCACLCH1, comprising =60 kDa of the anti-carcinoma monoclonal antibody (mAb) CC49. The single-chain protein consisted of the heavy-and light-chain variable (VH and VL) domains of the mAb covalently joined through a short linker peptide, while the carboxyl end of the VL domain was linked to the amino terminus of the human yl Fc reglon through the hinge region.The single-chain protein assembled into a dimeric molecule, termed SCAACLCH1, of 4120 kDa and was secreted into the tissue culture fluid. SDS/PAGE analysis of the secreted immunoglobulin purified by protein G affinity chromatography Monoclonal antibody (mAb) CC49, a murine IgGl (1), is a second-generation monoclonal of mAb B72.3 (2), which reacts with the tumor-associated glycoprotein TAG-72 (3) expressed on a variety ofcarcinomas. Murine mAb CC49 was developed by immunizing mice with TAG-72 purified by B72.3 affinity chromatography. Compared with B72.3, mAb CC49 has a higher antigen binding affinity (1) and targets human colon carcinoma xenografts in mice more efficiently and reduces the growth of the xenograft with greater efficacy (4, 5). Chimeric B72.3 with a human yl constant region has been shown to efficiently mediate antibody-dependent cellular cytotoxicity (ADCC). Results from ongoing clinical trials suggest that murine mAb CC49 is potentially a useful clinical reagent for targeting human colorectal carcinoma lesions.More recently, a single-chain Fv (sFv) fragment derived from the murine mAb CC49 was found to have an extremely rapid plasma and whole-body clearance rate in mice and rhesus monkeys (6). The sFv showed more rapid tumor penetration and more even distribution throughout the tumor mass (7). Recent studies have also shown, however

show abstract

“…Binding of cellular immune complexes to Fc recep tors induces cytotoxic events, among them 'antibody-de pendent cellular cytotoxicity'. This leads to activation of the effector cells and mediator release, probably an im portant process involved in specific tumor cell killing [9][10][11][12]. Therefore we focused our interest on the in situ distri bution of immunoglobulins in tumors as compared to con trol tissues.…”

mentioning

confidence: 99%

Distribution of Immunoglobulins in Squamous Cell Carcinoma of the Head and Neck

Neuchrist¹,

Kornfehl

Grasl³

et al. 1994

Int Arch Allergy Immunol

View full text Add to dashboard Cite

The immune response with respect to immunoglobulin production in the tumor was investigated in 23 patients with advanced squamous cell carcinomas of the head and neck. Immunohistochemical staining with monoclonal antibodies against IgG, IgM, IgA, IgD and IgE in the tumor was compared to normal hypopharyngeal mucosa. For IgG, IgA and IgM no significant differences between tumor and control tissues could be found. In contrast, a high number of IgE-positive cells was counted in most squamous cell carcinomas compared to normal mucosa. Most of these cells appeared as plasma cells. Regarding IgD the differences between tumor and control tissues, were less pronounced but also significant.

show abstract

Chimeric B72.3 mouse/human (IgG1) antibody directs the lysis of tumor cells by lymphokine-activated killer cells

Cited by 10 publications

References 37 publications

Transient expression of a tumor-specific single-chain fragment and a chimeric antibody in tobacco leaves

Transient expression of a tumor-specific single-chain fragment and a chimeric antibody in tobacco leaves

Secretion of a single-gene-encoded immunoglobulin from myeloma cells.

Distribution of Immunoglobulins in Squamous Cell Carcinoma of the Head and Neck

Contact Info

Product

Resources

About