Development of specific monoclonal antibodies and a sensitive discriminatory immunoassay for the circulating tumor markers SCCA1 and SCCA2

Çataltepe, Sule; Schick, Charles; Luke, Cliff J.; Pak, Stephen C.; Goldfarb, Daniel; Chen, Philip; Tanasiyevic, Milenko J.; Posner, Marshall R.; Silverman, Gary A.

doi:10.1016/s0009-8981(00)00197-2

Cited by 41 publications

(39 citation statements)

References 20 publications

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“…2). This finding is compatible with the earlier findings that SCCA2 is relatively underestimated when using the Abbott IMx system [21,23]. The workshop antibodies allow the establishment of several new assays for various forms of SCC.…”

Section: Discussionsupporting

confidence: 91%

“…When using differentiating antibodies, both SCCA1 and SCCA2 were expressed in the suprabasal layers of the stratified squamous epithelium of the normal uterine cervix and no differential expression between these SCCA in normal or malignant tissues was seen [21]. When using RT-PCR, however, the relative expression of SCCA2 over SCCA1 was increased in cervical cancer [22,23].…”

Section: Introductionmentioning

confidence: 94%

“…New methods were recently developed to measure serum SCCA1 or SCCA2 separately. By measuring recombinant SCCA preparations, it was demonstrated that rSCCA2 is underestimated by using the IMx system with a factor 1.5 or 2.5 [21,23].…”

Section: Introductionmentioning

confidence: 99%

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Characterization of Monoclonal Antibodies Directed against Squamous Cell Carcinoma Antigens: Report of the TD-10 Workshop

Nustad¹,

Dowell²,

Davis³

et al. 2004

Hum Hered

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Thirteen monoclonal antibodies directed against squamous cell carcinoma antigens (SCCA1 and SCCA2) were obtained from five international collaborating laboratories participating in the ISOBM TD-10 Workshop. Native and recombinant forms of SCCA were used in a wide variety of approaches to determine the reactivity and specificity of these antibodies. Based on reactivity, the antibodies could be divided into three groups: the SCCA1-reactive group containing those that reacted only with recombinant SCCA1 (rSCCA1) and native SCCA1 (nSCCA1) antigens, the SCCA2-reactive group containing those that reacted only with recombinant SCCA2 (rSCCA2), and the pan-reactive group containing those antibodies that reacted with rSCCA1, nSCCA1, and rSCCA2. Binding to radioiodinated rSCCA1 showed that all reactive antibodies were of a high affinity (K_d <2 × 10^–9 mol/l). Binding to labelled rSCCA2 demonstrated that five antibodies were of a high affinity (K_d <2 × 10^–9 mol/l). Antibody reactivity on Western blots was tested with nonreduced and reduced native and recombinant SCCA1 and SCCA2. In general, these findings showed that reduction had little effect on binding to SCCA1, but often a strong effect on the binding to SCCA2. Binding of antibodies to rSCCA1 and rSCCA2 in complexes with cathepsin L and G, respectively, was used to assist in the localization of epitope regions in enzyme-complexed SCCA. Cross-inhibition experiments showed that SCCA1-reactive antibodies represent two different epitope groups, and this is supported by their ability to make SCCA1-specific assays by combining antibodies from the two epitope groups. The SCCA2-reactive group represents two related antibodies and one unique as seen in cross-inhibition, but they do not form successful assay combinations. Classification of the pan-reactive antibodies is more difficult, as some epitope groups differ when results from rSCCA1 are compared with rSCCA2 as the target. However, two antibodies are outstanding, SCC107 and SCC113, as they are high-affinity antibodies which react equally well with free and protease complexes of SCCA1 and SCCA2. The precise location of epitopes was further studied using sequential overlapping peptides and homology modelling. The findings from this workshop strongly indicate that the recombinant antigens (rSCCA1 and rSCCA2) are very similar in epitope structure to the native counterparts in saliva, and squamous epithelium from normal and cancer tissues. Therefore, it is reasonable to conclude that the specificities found are reliable and have application for antibody measurement of all forms of squamous cell carcinoma in serum except SCCA2 in complex with its protease.

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Section: Discussionsupporting

confidence: 91%

Section: Introductionmentioning

confidence: 94%

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Characterization of Monoclonal Antibodies Directed against Squamous Cell Carcinoma Antigens: Report of the TD-10 Workshop

Nustad¹,

Dowell²,

Davis³

et al. 2004

Hum Hered

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“…Ç ataltepe et al 29 demonstrated that SCCA1 was elevated in amniotic fluids compared with SCCA2. To the best of our knowledge, this is the first report showing predominant elevation of SCCA2 in patients with diseases other than cancers.…”

Section: Discussionmentioning

confidence: 99%

The usefulness of combined measurements of squamous cell carcinoma antigens 1 and 2 in diagnosing atopic dermatitis

Shibata

Nakao

Azuma³

et al. 2012

Ann Clin Biochem

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Background: The squamous cell carcinoma antigen (SCCA) is widely used as a serological biomarker for various cancers. There are two known SCCA molecules, SCCA1 and SCCA2. We previously found that interleukin-4 or interleukin-13, two related Th2-type cytokines that play an important role in allergic diseases, induce expression of SCCA1 and SCCA2. In this study, we examined whether combined measurements of SCCA1 and SCCA2 are useful for diagnosing atopic dermatitis (AD). Methods: We established new enzyme-linked immunosorbent assays (ELISAs) to specifically detect SCCA1 or SCCA2. We applied serum samples from AD patients with food allergies and from cervical cancer patients to these ELISAs. We performed receiver operating characteristic analyses to diagnose AD and to distinguish AD from cervical cancer. Results: Serum concentrations of both SCCA1 and SCCA2 were elevated in AD patients. The serum concentrations of SCCA1 and SCCA2 positively correlated with the clinical severity of AD, showing high specificity (0.86 -0.88) and sensitivity (0.86) against control donors. The serum concentrations of SCCA1 and SCCA2 were elevated in cervical cancer patients; however, the SCCA2/SCCA1 ratios clearly distinguished AD patients from cervical cancer patients with high specificity (0.87) and sensitivity (0.87). Expression of SCCA2 was predominant in AD patients, whereas cervical cancer patients showed a predominance of SCCA1. Conclusions: Combined measurements of SCCA1 and SCCA2 are very useful in estimating the severity of allergic diseases, making it possible to distinguish allergic diseases from cancers.

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“…Immunoblotting was performed under reducing conditions as previously described (32). The protein bands were visualized by chemiluminescence (ECL Western Blotting Analysis System; Amersham Biosciences, Piscataway, NJ), and quantitated by densitometry with NIH-Image 163.…”

Section: Immunoblottingmentioning

confidence: 99%

Imbalance between Cysteine Proteases and Inhibitors in a Baboon Model of Bronchopulmonary Dysplasia

Altiok

Yasumatsu

Bingol-Karakoc

et al. 2006

Am J Respir Crit Care Med

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Development of specific monoclonal antibodies and a sensitive discriminatory immunoassay for the circulating tumor markers SCCA1 and SCCA2

Cited by 41 publications

References 20 publications

Characterization of Monoclonal Antibodies Directed against Squamous Cell Carcinoma Antigens: Report of the TD-10 Workshop

Characterization of Monoclonal Antibodies Directed against Squamous Cell Carcinoma Antigens: Report of the TD-10 Workshop

The usefulness of combined measurements of squamous cell carcinoma antigens 1 and 2 in diagnosing atopic dermatitis

Imbalance between Cysteine Proteases and Inhibitors in a Baboon Model of Bronchopulmonary Dysplasia

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