Single-Chain Fv Fusion Proteins Suitable as Coating and Detecting Reagents in a Double Antibody Sandwich Enzyme-Linked Immunosorbent Assay

1997 ◽  
Vol 249 (2) ◽  
pp. 219-227 ◽  
Author(s):  
Randolf J. Kerschbaumer ◽  
Sonja Hirschl ◽  
Andrea Kaufmann ◽  
Martin Ibl ◽  
Renate Koenig ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Fusion Proteins ◽  
Enzyme Linked Immunosorbent Assay ◽  
Single Chain ◽  
Double Antibody ◽  
Single Chain Fv

A double antibody sandwich enzyme-linked immunosorbent assay for detection of secreted antigen 1 of Babesia microti using hamster model

2012 ◽  
Vol 130 (2) ◽  
pp. 178-182 ◽  
Author(s):  
Yuzi Luo ◽  
Mohamad Alaa Terkawi ◽  
Honglin Jia ◽  
Gabriel Oluga Aboge ◽  
Youn-Kyoung Goo ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Babesia Microti ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hamster Model ◽  
Double Antibody

A modified double antibody sandwich enzyme-linked immunosorbent assay for measurement of alpha-1-antitrypsin in biologic fluids

1985 ◽  
Vol 83 (1) ◽  
pp. 101-112 ◽  
Author(s):  
Joseph P. Michalski ◽  
Candace C. McCombs ◽  
Sangita Sheth ◽  
Maria McCarthy ◽  
Richard deShazo
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Double Antibody ◽  
Alpha 1 Antitrypsin

Immuno-enzymatic assay for the detection of Schistosoma mansoni antigens in serum of mice harbouring bisexual or unisexual light worm infections

1979 ◽  
Vol 53 (3) ◽  
pp. 189-194 ◽  
Author(s):  
A. W. Ferreira ◽  
A. L. M. Caldini ◽  
S. Hoshino-Shimizu ◽  
M. E. Camargo
Keyword(s):  
Schistosoma Mansoni ◽  
Immunosorbent Assay ◽  
Enzymatic Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Antigens ◽  
Male Worm ◽  
Antibody Technique ◽  
Post Exposure ◽  
Double Antibody

ABSTRACTEnzyme-linked immunosorbent assay (ELISA) double antibody technique was standardized for detecting S. mansoni Polysaccharide and protein antigens in serum of infected mice. Anti-sera specific for either worm components were obtained in sheep and peroxidase conjugates prepared from each serum. The immunoenzimatic test for protein could detect as little as 6μg/ml antigens, and the test for polysaccharides 3 μg/ml.Both bisexual and unisexual male worm low infections were produced, and studied for as long as 27 weeks post-exposure. Worm components were found in serum from both types of infections and in progressively higher percentages of animals until the end of the 27 weeks observations period. For unisexual male infections this percentage reached from about 50% to 60% of mice, and 100% for bisexual infections. Significantly higher antigen concentrations in serum were found at 27 weeks for bisexual infections, no antigen increase being detected in relation to starting egg secretion, which occurred at 5 week infections.

A vaccine approach for breast cancer immunotherapy by targeting HER2/neu antigen to B cells via CD19 molecule.

2011 ◽  
Vol 29 (27_suppl) ◽  
pp. 287-287
Author(s):  
D. Xiang ◽  
Y. Ma ◽  
C. Ding ◽  
G. H. Kloecker ◽  
J. Yan
Keyword(s):  
Breast Cancer ◽  
B Cells ◽  
Fusion Protein ◽  
Tumor Cells ◽  
Fusion Proteins ◽  
In Vivo Studies ◽  
Single Chain ◽  
Tumor Vaccines ◽  
Single Chain Fv ◽  
Her 2

287 Background: Trastuzumab has been proven to be effective in the immunotherapy for Her-2/neu-positive breast cancer. However, this adoptively transferred therapeutic Ab must be continuously given to the patient with huge financial cost. Thus, it would be desirable if tumor vaccines could elicit long-lasting trastuzumab-like Ab. Methods: Constructing fusion protein of anti-mouse CD19 single chain variable fragment (anti-CD19-scFv) with human Her-2/neu extracellular domain P3-4 (P3-4) by extracting total RNA from Rat anti-mouse CD19 hybridoma (1D3) and first strand cDNA was synthesized accordingly. VH and VL were amplified using designed primers. Single chain Fv (VL-VH) was then synthesized by overlapping PCR. Then constructs were sequenced and cloned into pET-20b(+) vectors. Fragments P3-4 of extracellular domains of human Her-2/neu was cloned from pcDNA3.1-Her2 and then cloned into pET-20b(+)-scFv. Series of constructs were expressed and purified according to standard protocol and verified by Western Blot. Results: Both in vitro and in vivo studies demonstrated that fusion proteins anti-CD19-scFv and anti-CD19-scFv-P3-4 but not P3-4 could specifically bind to B cells with high affinity. Mice immunized with anti-CD19-scFv-P3-4 secreted higher titers of IgG and IgM than those from controls (p<0.05). Studies also demonstrated that sera from anti-CD19-scFv-P3-4 but not anti-CD19-scFv or P3-4 immunized mice stained with Her-2/neu expressing SKOV-3 tumor cells. These Abs also competitively inhibited trastuzumab-mediated Ag binding, suggesting that trastuzumab-like Ab responses were elicited. WT mice immunized with anti-CD19-scFv-P3-4 fusion protein then challenged with D2/F2-Her-2 mammary tumor cells showed significantly reduced tumor burden compared to those immunized with control fusion proteins (p<0.05) and had enhanced median overall survival to 45 days versus 34 days in WT mice immunized with either anti-CD19-scFv or P3-4. Conclusions: Targeting of Her-2/neu antigens to B cells stimulates Th-dependent humoral immune responses with anti tumor effect in mouse model. These findings provide a novel avenue for successful development of breast cancer vaccination strategy and help to fight for cancer.

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