Kinetic and affinity constants of epitope specific anti-carcinoembryonic antigen (CEA) monoclonal antibodies for CEA and engineered CEA domain constructs

Hefta, Laura J. F.; Neumaier, Michael; Shively, John E.

doi:10.1016/s1380-2933(98)00004-9

Cited by 42 publications

(19 citation statements)

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“…The CEA-specific mAb T84.66 has a very high affinity constant (K D ϭ 8 pM) and does not crossreact with other members of the CEA family (26,43). These properties make this antibody an ideal candidate for tumor detection and immunotherapy.…”

Section: Discussionmentioning

confidence: 99%

“…Because CEA can be detected in almost all human colon cancers, 50% of all breast cancers, and in other tumors of epithelial origin, anti-CEA antibodies have been used for antibody-mediated cancer therapies and in vivo tumor imaging. Among those, the mAb T84.66, which binds to the A3 domain of CEA with high specificity and affinity (K D ϭ 8 pM) (26), has been used successfully for in vivo imaging and diagnosis of human colorectal carcinoma (27). A recombinant mouse͞human chimeric antibody (cT84.66), a minibody (scFv-CH3), and a scFv fragment (scFvT84.66) recently have been engineered, and the expressed proteins were characterized and evaluated for diagnostic and therapeutic applications (28)(29)(30)(31)(32).…”

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confidence: 99%

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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

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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...

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Section: Discussionmentioning

confidence: 99%

mentioning

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

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“…CEA, originally identified in human fetal intestine and adult CC tissue in 1965 by Gold and Freedman (6), was subsequently characterized as a glycosylated cell-surface glycoprotein with a molecular weight of 180,000 Da (7); further research identified it as a significant tumor-associated antigen that is highly overexpressed in breast, lung and pancreatic cancer and, particularly, in CRC (8,9). CEA is routinely detected as a tumor biomarker and an auxiliary indicator for the preoperative diagnosis of CRC (10), as well as an early predictor of recurrence (5), which to some extent accounts for CEA reflecting the nature of CRC.…”

Section: Introductionmentioning

confidence: 99%

Differences in carcinoembryonic antigen levels between colon and rectal cancer

Ding

Xuan

Chen

et al. 2014

Molecular and Clinical Oncology

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“…Human/murine chimeric T84.66, an anti-CEA intact IgG 1 with high affinity (affinity constant ϭ 1 ϫ 10 11 M Ϫ1 ) 17 and specificity for CEA, was prepared according to published methods, 18 and labeled with 124 I, using the iodogen method. 19 Iodine-124 was obtained as NaI (radiochemical purity greater than 95%, radionuclidic purity greater than 99.9% at initial calibration) from IBA/Eastern Isotopes (Sterling, VA).…”

Section: Radiotracersmentioning

confidence: 99%

Quantitative Serial Imaging of an ¹²⁴I Anti-CEA Monoclonal Antibody in Tumor-Bearing Mice

Bading

Hörling

Williams

et al. 2008

Cancer Biotherapy and Radiopharmaceuticals

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Kinetic and affinity constants of epitope specific anti-carcinoembryonic antigen (CEA) monoclonal antibodies for CEA and engineered CEA domain constructs

Cited by 42 publications

References 4 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

Differences in carcinoembryonic antigen levels between colon and rectal cancer

Quantitative Serial Imaging of an ¹²⁴I Anti-CEA Monoclonal Antibody in Tumor-Bearing Mice

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Kinetic and affinity constants of epitope specific anti-carcinoembryonic antigen (CEA) monoclonal antibodies for CEA and engineered CEA domain constructs

Cited by 42 publications

References 4 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

Differences in carcinoembryonic antigen levels between colon and rectal cancer

Quantitative Serial Imaging of an 124I Anti-CEA Monoclonal Antibody in Tumor-Bearing Mice

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Quantitative Serial Imaging of an ¹²⁴I Anti-CEA Monoclonal Antibody in Tumor-Bearing Mice