Cell surface receptor-antibody association constants and enumeration of receptor sites for monoclonal antibodies

Siiman, Olavi; Burshteyn, Alexander

doi:10.1002/1097-0320(20000801)40:4<316::aid-cyto7>3.0.co;2-c

Cited by 25 publications

(27 citation statements)

References 28 publications

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“…Ag particle-Amdex colloidal suspensions 1.215 milliliters of 0.589 mol/dm 3 silver nitrate solution were added to 95 ml of DI water in a 250-ml flask. The contents of the flask were then heated to about 90°C, upon which 10 ml of a 40 mg/ml solution of 5X-Amdex were added to the reaction flask.…”

Section: Amdex-3mentioning

confidence: 65%

Surface‐enhanced Raman scattering (SERS) of random silver or gold particle arrays on aminodextran‐coated polystyrene beads

Siiman¹,

Ledis²

2005

J Raman Spectroscopy

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Citrate and/or chemically modified aminodextran (Amdex) SERS bands were detected from silver particleAmdex-polystyrene (PS) beads either as singlets or small clusters, prepared by reaction between aqueous silver nitrate, Amdex-coated PS beads, and aqueous sodium citrate solution. However, citrate on individual silver or gold particles of the same size was not detected with 633-nm excitation. SERS of dyes such as rhodamine 6 G was readily observed on individual silver particles or on the gold beads. Three distinct modes of citrate/modified-Amdex binding to the silver particle-Amdex-PS beads were characterized: (1) exchangeable citrate bound through one or more carboxylate groups -citrate could be washed away with H 2 O or D 2 O; displaced by successively greater concentrations of R6G; or squeezed out mechanically by applying the pressure of a second coverslip on top of a droplet of bead suspension; (2) nonexchangeable Amdex bound to silver via a primary alcoholate ion of oxidized, glutaraldehyde-cross-linked Amdex; and (3) nonexchangeable oxidized Amdex bound to silver via carboxylate group(s) but with H/D exchangeable alcohol hydrogen atom. That oxidation of the glucopyranose residues of Amdex on PS beads had taken place by Ag(I) as silver nitrate in aqueous solution to split out carboxylate groups was first identified from their characteristic vibrational modes in the SERS spectra of colloidal silver suspensions.Minimum amplification factors for (R6G on Ag)/(R6G on Au) with 633-nm excitation were estimated as 1 × 10 15 /1 × 10 11 . An extra SERS enhancement of about 1000-fold due to multiplicity of metal particles per bead was included.

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Section: Amdex-3mentioning

confidence: 65%

Surface‐enhanced Raman scattering (SERS) of random silver or gold particle arrays on aminodextran‐coated polystyrene beads

Siiman¹,

Ledis²

2005

J Raman Spectroscopy

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“…The binding is non-covalent with an association constant of 10 15 M -1 (Diamandis & Christopoulos 1991), and is therefore stronger than the association constant when an antibody binds to its receptor, reported to be in the range of 10 8 -10 12 M -1 (Siiman & Burshteyn 2000, Xie et al 2005. The binding forms rapidly and is convenient for proof-of-concept studies as those performed in paper I and II.…”

Section: Bioconjugation Of the Targeting Ligand And The Protein-toxinmentioning

confidence: 99%

Photochemically stimulated drug delivery increases the cytotoxicity and specificity of EGF–saporin

Weyergang

Selbo

Berg

2006

Journal of Controlled Release

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“…This high-molecular-weight band is probably the Bcl1 protein, known as the most immunogenic antigen of the exosporium fraction (14,20,22). The affinity constant of the purified scFv antibody was determined via FCM as demonstrated previously (18,27). In short, mean spore fluorescence at steady state was measured as a function of the antibody concentration.…”

Section: Resultsmentioning

confidence: 99%

“…The affinity of the purified scFv antibody for B. anthracis spores was determined using flow cytometry (FCM) on a FACSCalibur machine (Becton Dickinson Immuno Cytometry Systems, Mississauga, Ontario, Canada), as described previously (18,27). Each test tube contained B. anthracis spores (1 ϫ 10 6 CFU/ml) and anti-E-tag (Pharmacia) and fluorescein isothiocyanate (FITC)-conjugated anti-mouse (F2266; Sigma-Aldrich, St. Louis, MO) (2 g/ml) antibodies.…”

Section: Bacteria B Anthracis ⌬14185 Is a Nontoxinogenic Nonencapsmentioning

confidence: 99%

Development and Implementation of a Single-Chain Fv Antibody for Specific Detection of Bacillus anthracis Spores

Mechaly

Zahavy

Fisher

2008

Appl Environ Microbiol

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A single-chain Fv (scFv) antibody was developed and applied for efficient and specific detection of Bacillus anthracis spores. The antibody was isolated from a phage display library prepared from spleens of mice immunized with a water-soluble extract of the outer membrane of the B. anthracis spore (exosporium). The library (7 ؋ 10 6 PFU) was biopanned against live, native B. anthracis ATCC ⌬14185 spores suspended in solution, resulting in the isolation of a unique soluble scFv antibody. The antibody was affinity purified and its affinity constant (3 ؋ 10 8 ؎ 1 ؋ 10 8 M ؊1 ) determined via flow cytometry (FCM). Preliminary characterization of scFv specificity indicated that the scFv antibody does not cross-react with representatives of some phylogenetically related Bacillus spores. The potential use of scFv antibodies in detection platforms was demonstrated by the successful application of the soluble purified scFv antibody in enzyme-linked immunosorbent assays, immunofluorescence assays, and FCM.Bacillus anthracis spores, the primary infectious agents causing anthrax, are probably the most likely candidates for a biological terrorist assault. Therefore, rapid detection of spores is critical for a timely response and successful treatment of the disease. Various immunoassays based on the high specificity of antibodies have been developed for the rapid detection of several pathogens. Over the past decade, recombinant technology enabled the production of engineered antibody fragments such as Fabs or single-chain Fv (scFv) antibodies. An scFv antibody is a small engineered antibody in which the variable heavy chain and light chain of the antibody molecule are connected by a short, flexible polypeptide linker. Phage display technology enables the presentation of scFv antibody on the phage surface and has been used successfully for the isolation of specific scFv antibodies from animal repertoire libraries via several enrichment cycles.Using scFv antibodies for antigen detection has several advantages. scFv antibodies can be produced in large quantities in bacterial expression systems, with high reproducibility at low cost, and can be manipulated genetically for improved specificity and affinity (5,15,17). The recombinant antibody can also be fused to marker molecules for detection purposes (25). Recombinant antibodies have been developed for treatment of anthrax infection (26) and disease detection in clinical applications (25). However, the use of recombinant antibodies for detection of B. anthracis spores has not been described. In this study, we describe the construction of an scFv antibody and its successful application for B. anthracis spore detection. Bacillus cultures and sporulation. Spores of all strains were produced in SSM sporulation medium, as previously described (6). MATERIALS AND METHODS Bacteria. B. anthracis ⌬14185 is a nontoxinogenic, nonencapsulated (ToxImmunization. Female BALB/c mice were immunized subcutaneously with 1 ϫ 10 7 CFU of irradiated (15 min at maximum intensity in a microwave oven) B. ...

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Cell surface receptor-antibody association constants and enumeration of receptor sites for monoclonal antibodies

Cited by 25 publications

References 28 publications

Surface‐enhanced Raman scattering (SERS) of random silver or gold particle arrays on aminodextran‐coated polystyrene beads

Surface‐enhanced Raman scattering (SERS) of random silver or gold particle arrays on aminodextran‐coated polystyrene beads

Photochemically stimulated drug delivery increases the cytotoxicity and specificity of EGF–saporin

Development and Implementation of a Single-Chain Fv Antibody for Specific Detection of Bacillus anthracis Spores

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