The main goal of this competitive renewal application is the full understanding of the nature of breast epithelial antigens (BEAgs) found in the circulation of breast tumors and the use of this knowledge to develop the most sensitive and specific assays for breast cancer diagnosis. Emphasis will be placed on identifying molecular and epitopic structures and maturational characteristics of BEAgs in breast cancer patient serum, so as to be able to synthesize novel recombinant epitopes, both glycosylated or not, of anti-BEAgs to participate in highly sensitive competitive recombinant epitope assays (REAs) for these breast cancer serum markers. Several of these recombinant epitopes will be expressed in fusion proteins that will participate as solid phase-incorporated antigen in REA panels. For these REAs the affinity of the participating antibody will be similar or lower for the solid phase-incorporated antigen than for competing serum breast cancer antigen glycoforms. Panels of these REAs will be created that will incorporate epitopes of BEAgs (the breast epithelial mucins, the 46 kDa Ag and conventional ones such as CEA). Sensitivity of these REAs will be heightened by protein design and molecular engineering to increase the affinity of the anti- BEAgs participating in them. This technology is represented by phage display libraries employing computer guided mutagenesis of the CDRs and by immunoglobulin chain shuffling in filamentous phage. Crucial to increasing the sensitivity of REAs is the ability acquired through this project using molecular design, to control the interplay between solid phase-incorporated fusion protein, the affinity- increased antibody and the competing breast serum antigen. The highest possibly sensitivity, while preserving the high specificity already demonstrated by the REAs, will be required to measure accurately the low levels of BEAgs to be found in the targeted subgroups potentially bearing small breast tumors usually difficult to detect: the axillary-node free patient with residual disease and the under 50 year old women, whose mammographic diagnosis is now in question. These molecularly redesigned and optimized REAs will participate as a second diagnostic test to mammograms. Their combined ability to detect small breast tumors will be determined and then compared to that of either test alone. A new area of exploration for this project will be the use of piezoelectric microsensors to increase sensitivity of detection of REAs. The advanced methodology for immunoassay optimization proposed in this project could represent a model that could have impact in the general area of serological assay development.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA039932-13
Application #
2458039
Study Section
Pathology B Study Section (PTHB)
Program Officer
Lively, Tracy (LUGO)
Project Start
1984-09-16
Project End
1999-07-31
Budget Start
1997-08-01
Budget End
1999-07-31
Support Year
13
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Cancer Research Fund of Contra Costa
Department
Type
DUNS #
City
Walnut Creek
State
CA
Country
United States
Zip Code
94598
Peterson, J A; Scallan, C D; Ceriani, R L et al. (2001) Structural and functional aspects of three major glycoproteins of the human milk fat globule membrane. Adv Exp Med Biol 501:179-87
Peterson, J A; Hamosh, M; Scallan, C D et al. (1998) Milk fat globule glycoproteins in human milk and in gastric aspirates of mother's milk-fed preterm infants. Pediatr Res 44:499-506
Newburg, D S; Peterson, J A; Ruiz-Palacios, G M et al. (1998) Role of human-milk lactadherin in protection against symptomatic rotavirus infection. Lancet 351:1160-4
Peterson, J A; Blank, E W; Ceriani, R L (1997) Effect of multiple, repeated doses of radioimmunotherapy on target antigen expression (breast MUC-1 mucin) in breast carcinomas. Cancer Res 57:1103-8
DeNardo, S J; Kramer, E L; O'Donnell, R T et al. (1997) Radioimmunotherapy for breast cancer using indium-111/yttrium-90 BrE-3: results of a phase I clinical trial. J Nucl Med 38:1180-5
Couto, J R; Taylor, M R; Godwin, S G et al. (1996) Cloning and sequence analysis of human breast epithelial antigen BA46 reveals an RGD cell adhesion sequence presented on an epidermal growth factor-like domain. DNA Cell Biol 15:281-6
Peterson, J A; Couto, J R; Taylor, M R et al. (1995) Selection of tumor-specific epitopes on target antigens for radioimmunotherapy of breast cancer. Cancer Res 55:5847s-5851s
Ceriani, R L; Blank, E W; Couto, J R et al. (1995) Biological activity of two humanized antibodies against two different breast cancer antigens and comparison to their original murine forms. Cancer Res 55:5852s-5856s
Ortel, T L; Quinn-Allen, M A; Keller, F G et al. (1994) Localization of functionally important epitopes within the second C-type domain of coagulation factor V using recombinant chimeras. J Biol Chem 269:15898-905
Peterson, J A; Ceriani, R L (1994) Breast mucin and associated antigens in diagnosis and therapy. Adv Exp Med Biol 353:1-8

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