Targeting the proteome/epitome, implementation of subtractive immunization

Zijlstra, Andries; Testa, Jacqueline E.; Quigley, James P.

doi:10.1016/s0006-291x(03)00357-7

Cited by 23 publications

(18 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To produce monoclonal antibodies specific for pancreatic juice from PC patients, we immunized mice using subtractive immunization. This method is useful to produce antibodies specific to the antigen that are poorly immunogenic or have low abundance of the immunogen (10)(11)(12). Indeed, in this study, post-immunized mouse serum showed immunogenspecific bands that were different from CA19-9 and CEA, despite an abundance of these antigens in both the tolerogen and immunogen.…”

Section: Discussionmentioning

confidence: 67%

“…Thus, we used a subtractive immunization method. Subtractive immunization is an efficient way to produce antibodies against poorly immunogenic or rare antigens (10)(11)(12). Mice were tolerized against the tolerogen using the immunosuppressive drug cyclophosphamide (Cy).…”

Section: Introductionmentioning

confidence: 99%

“…Mice were tolerized against the tolerogen using the immunosuppressive drug cyclophosphamide (Cy). Cy preferentially eliminates T and B cells activated by the tolerogen (11,13). After tolerization, mice were immunized with the immunogen.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Novel monoclonal antibodies against pancreatic juice from pancreatic cancer patients and their possible application in differential diagnosis

Hamabashiri

Daichou²,

Yasumoto³

et al. 2011

Int J Mol Med

View full text Add to dashboard Cite

Abstract. Pancreatic cancer (PC) has a poor clinical prognosis with a <10% 5-year survival rate. Because there are no specific biomarkers of PC, it is difficult to detect small PC tumors and most patients are diagnosed at an advanced stage. Specific biomarkers are useful tools for the early detection of cancer. However, PC-related biomarkers, such as CA19-9 lack specificity and sensitivity. In this study, we took an immunological approach to establish novel monoclonal antibodies (mAbs) specific for the pancreatic juice from PC patients, which would be potentially useful in the diagnosis of PC. Mice were immunized by subtractive immunization using mixed pancreatic juices from chronic pancreatitis and PC patients as the tolerogen and the immunogen, respectively. After screening by Western blotting, four mAbs were obtained: 2P-1-2-1, 2P-1-17-1, 6P-3-2-4 and 7P-9-11-6. The mAb 2P-1-2-1 showed reactivity against the tolerogen at 115 and 120 kDa, but only the 120-kDa antigen was also reactive to the immunogen. The mAb 2P-1-17-1 showed an intense smear reactivity at ~150 kDa against the immunogen. Finally, the mAbs 6P-3-2-4 and 7P-9-11-6 showed PC-specific reactivity to the immunogen at >250 kDa and at ~70 kDa, respectively. We propose that investigation of pancreatic juice samples with these mAbs may enable us to perform reliable differential diagnosis of benign and malignant diseases. Furthermore, we demonstrated that subtractive immunization is a useful method for producing mAbs specific for the pancreatic juice from PC patients.

show abstract

Section: Discussionmentioning

confidence: 67%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Novel monoclonal antibodies against pancreatic juice from pancreatic cancer patients and their possible application in differential diagnosis

Hamabashiri

Daichou²,

Yasumoto³

et al. 2011

Int J Mol Med

View full text Add to dashboard Cite

show abstract

“…This problem is potentially exacerbated by the weak immunogenicity of some tumor antigens. The impact of dominant epitopes can be minimized by using a subtractive immunization approach (Hooper et al 2003, Zijlstra et al 2003 or by incorporating a highly selective screen.…”

Section: Monoclonal Antibodiesmentioning

confidence: 99%

Identification and validation of cell surface antigens for antibody targeting in oncology

Carter

Smith²,

Ryan³

2004

Endocr Relat Cancer

109

View full text Add to dashboard Cite

Recent clinical successes with antibodies have reinvigorated interest in the identification and validation of new antigens for antibody therapy, including cell surface proteins for targeting in oncology, the focus of this review. Target identification commonly involves the search for differences between tumor and non-tumor cell lines and/or tissue at the DNA, mRNA, protein or antibody reactivity levels. The next stage, target validation, utilizes antibodies to profile the expression of antigen in normal and tumor tissue and to verify that the antigen is selectively expressed on the surface of tumor cells. Supportive evidence for protein expression is often sought by mRNA profiling and, sometimes, analysis for genomic defects. Unfortunately, concordance between mRNA and protein levels has been found in only about $20% of cases and therefore must be evaluated for individual targets of interest. Antigens judged suitable for antibody targeting are then advanced to the next stage, namely, in vitro and then in vivo screening of antibodies for anti-tumor activities. Subsequent optimization of an antibody clinical lead for therapy is a desirable, if not obligatory, step to developing an antibody as an anti-cancer therapeutic. No single approach or even combination of methods has emerged as the preferred way to identify surface antigens suitable for targeting in oncology. Major options at each step in the process are reviewed here, including their strengths and limitations.

show abstract

“…Nonetheless, epitope prediction remains a popular first screening method to identify candidate T cell determinants for subsequent biological validation (78 -89), and predictive algorithms are frequently combined with in vitro MHC-binding assays to confirm experimentally that the predicted ligands bind to the targeted MHC molecule (80,90). A more comprehensive approach that allows assessment of natural processing and presentation of candidate epitopes involves the direct biochemical analysis of class I or class II ligands, which has been coined as the immunoproteome (82,83,(91)(92)(93)(94)(95)(96)(97)(98)(99).…”

Section: Defining and Refining Mhc-binding Motifs And Their Use In Bimentioning

confidence: 99%

Immunoproteomics

Purcell

Gorman

2004

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

Targeting the proteome/epitome, implementation of subtractive immunization

Cited by 23 publications

References 59 publications

Novel monoclonal antibodies against pancreatic juice from pancreatic cancer patients and their possible application in differential diagnosis

Novel monoclonal antibodies against pancreatic juice from pancreatic cancer patients and their possible application in differential diagnosis

Identification and validation of cell surface antigens for antibody targeting in oncology

Immunoproteomics

Contact Info

Product

Resources

About