Preparation and characterization of monoclonal antibodies specific to synthetic peptide of carcinoembryonic antigen

Tsujisaki, Masayuki; Imai, Kohzoh; Hishikawa, Noriyuki; Tokuchi, Shigeru; Hinoda, Yuji; Matsukawa, Hirokazu; Oikawa, Shinzo; Nakazato, Hiroshi; Yachi, Akira

doi:10.1002/ijc.2910470216

Cited by 8 publications

(10 citation statements)

References 21 publications

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“…This approach has been successfully applied in our laboratory for generating antisera against the C-domain regions of two PSG products, i.e., PSGld and PSG3, and development of further antisera is in progress. Similarly, Tsujisaki et al (128) have raised antibodies against CEA peptides, some of which also recognize purified CEA.…”

Section: Identification Of Humanrodent Counterparts By Comparing Exprmentioning

confidence: 99%

Carcinoembryonic antigen gene family: Molecular biology and clinical perspectives

Thompson

Fritz

Zimmermann

1991

Clinical Laboratory Analysis

557

389

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The carcinoembryonic antigen (CEA) gene family belongs to the immunoglobulin supergene family and can be divided into two main subgroups based on sequence comparisons. In humans it is clustered on the long arm of chromosome 19 and consists of approximately 20 genes. The CEA subgroup genes code for CEA and its classical crossreacting antigens, which are mainly membrane-bound, whereas the other subgroup genes encode the pregnancy-specific glycoproteins (PSG), which are secreted. Splice variants of individual genes and differential post-translational modifications of the resulting proteins, e.g., by glycosylation, indicate a high complexity in the number of putative CEA-related molecules. So far, only a limited number of CEArelated antigens in humans have been unequivocally assigned to a specific gene.Rodent CEA-related genes reveal a high sequence divergence and, in part, a completely different domain organization than the human CEA gene family, making it difficult to determine individual gene counterparts. However, rodent CEA-related genes can be assigned to human subgroups based on similarity of expression patterns, which is characteristic for the subgroups. Various functions have been determined for members of the CEA subgroup in vitro, including cell adhesion, bacterial binding, an accessory role for collagen binding or ecto-ATPases activity.Based on all that is known so far on its biology, the clinical outlook for the CEA family has been reassessed.

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Section: Identification Of Humanrodent Counterparts By Comparing Exprmentioning

confidence: 99%

Carcinoembryonic antigen gene family: Molecular biology and clinical perspectives

Thompson

Fritz

Zimmermann

1991

Clinical Laboratory Analysis

557

389

View full text Add to dashboard Cite

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“…Tsujisaki et al [26] immunized mice with a 22-amino acid peptide (P1) corresponding to the beginning of the A1 domain (aa 119-140) and generated three Mabs. Two of them (P1-234 and P1-255) bound to peptide P1 and native CEA, whereas the third one (P1-706) only reacted with P1 and periodate-treated CEA.…”

Section: Discussionmentioning

confidence: 99%

“…Different anti-CEA MAbs have been developed following immunization with CEA-derived peptides [26][27][28]. Tsujisaki et al [26] immunized mice with a 22-amino acid peptide (P1) corresponding to the beginning of the A1 domain (aa 119-140) and generated three Mabs.…”

Section: Discussionmentioning

confidence: 99%

Carcinoembryonic Antigen Continuous Epitopes Determined by the SPOT Method

Solassol¹,

Granier²,

Pèlegrin³

2001

Tumor Biol

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Carcinoembryonic antigen (CEA) is a heavily glycosylated tumor-associated protein with an N-A1-B1-A2-B2-A3-B3 domain structure. Circulating CEA immunoassays are used for monitoring digestive cancer patients, and radiolabeled anti-CEA monoclonal antibodies (MAb) are used for the diagnosis and therapy of CEA-positive tumors. The five major nonoverlapping epitopes (Gold 1–5) have been broadly correlated with the domain organization, but there is no precise localization of the epitopes at the sequence level. In an attempt to identify the peptide sequences corresponding to the five Gold epitopes on the CEA molecule, we prepared a set of 227 overlapping fifteen-mer peptides corresponding to the complete CEA sequence with the SPOT method. Using five high affinity MAbs directed against the five CEA Gold epitopes, we demonstrated that none of these epitopes could be mimicked by a fifteen-mer peptide sequence. However, using rabbit and goat anti-CEA sera, we identified six major continuous antigenic regions. All are included in the Ig-like domains of the CEA: two in the A1 domain (residues 120–134 and 153–164), one each in the A2 (329–337) and A3 domains (508–513), one at the junction between the A3 and B3 domains (553–561) and one in the B3 domain (565–573). A very homologous sequence (common residues VSPRL) was mapped in each of the three A domains. Thus, in terms of occurrence of continuous epitopes, the Ig-like domains A1, A2, A3 and B3 seem to be the most antigenic parts of CEA. These peptide sequences should be good candidates for the future development of site-specific anti-CEA MAbs.

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“…MoAb P1-356 (IgG1) was prepared to a synthetic peptide (P1: 119-140 amino acids) in domain I of the CEA molecule, as described elsewhere (5). The MoAbs were secreted hybridomas constructed with myeloma cells and splenocytes from BALB/c mice, which were immunized once a week with three intraperitoneal injections of 100 mg of synthetic peptide P1 suspended in Freund's complete adjuvant.…”

Section: Development Of Moab To Synthetic Cea Peptide P1mentioning

confidence: 99%

“…In a previous study (5) we prepared MoAb P1-356 specific to synthetic peptide (P1: 119-140 amino acids) in domain I of carcinoembryonic antigen (CEA) as one of the main tumor markers of human colorectal carcinoma. In the present study, we prepared the anti-Id MoAbs (Ab2b) bearing an internal image of CEA, and the anti-anti-Id MoAb (Ab3), to investigate the internal image from the molecular aspect, and to determine what kind of anti-Id antibody is useful for vaccination or cancer therapy.…”

Section: Introductionmentioning

confidence: 99%

Preparation and characterization of anti‐anti‐idiotypic monoclonal antibody (“Ab1‐like Ab3”) in relation to carcinoembryonic antigen

Tsujisaki

Masuya

Okada

et al. 2002

Clinical Laboratory Analysis

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In order to analyze the epitope structure of carcinoembryonic antigen (CEA) and the idiotype network system, seven anti-idiotypic monoclonal antibodies (anti-Id MoAbs) were generated from a BALB/c mouse immunized with anti-CEA MoAb P1-356, which recognized a synthetic peptide P1 of CEA, domain I. These MoAbs were divided into four groups. The anti-Id MoAbs specifically reacted with MoAb P1-356, but not with any MoAb, and inhibited the binding of MoAb P1-356 to CEA, indicating that all of these anti-Id MoAbs recognized private idiotopes at the combining sites of MoAb P1-356. Polyclonal anti-anti-idiotypic antiserum (Ab3) generated with anti-Id MoAbs M315 (Ab2), blocked the binding of MoAb P1-356 (Ab1) to CEA and reacted with CEA(Ag) and synthetic peptide P1. The analysis of serological assays suggested that it contains "Ab1-like Ab3." Therefore, we prepared anti-anti-Id MoAbs using anti-Id MoAb M315. Among 13 candidates, anti-anti-Id MoAb 11B2 was selected, because it competed with MoAb P1-356 (Ab1) binding to CEA. A direct binding assay using MoAb11B2 showed that it reacted with purified CEA and with CEA synthetic peptide P1. In addition, MoAb 11B2 (Ab3) reacted with both CEA-producing cultured cells and colonic cancerous tissues in immunostaining. These results indicate that anti-anti-Id MoAb 11B2 is "Ab1-like Ab3." Therefore, it is suggested that anti-Id MoAb M315 bears an "internal image" of the MoAb P1-356-defined epitope on CEA.

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Preparation and characterization of monoclonal antibodies specific to synthetic peptide of carcinoembryonic antigen

Cited by 8 publications

References 21 publications

Carcinoembryonic antigen gene family: Molecular biology and clinical perspectives

Carcinoembryonic antigen gene family: Molecular biology and clinical perspectives

Carcinoembryonic Antigen Continuous Epitopes Determined by the SPOT Method

Preparation and characterization of anti‐anti‐idiotypic monoclonal antibody (“Ab1‐like Ab3”) in relation to carcinoembryonic antigen

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