Factors affecting the fine specificity and sensitivity of serum antiganglioside antibodies in ELISA

Ravindranath, Mepur H.; Ravindranath, Rajeswari M.H.; Morton, Donald L.; Graves, Michael C.

doi:10.1016/0022-1759(94)90270-4

Cited by 52 publications

(35 citation statements)

References 43 publications

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“…Antifouling agents such as polyethylene glycol can also be deposited on areas surrounding the sensor to prevent target depletion via nonspecific binding [62][63][64]. Use of blocking agents is not a systematic science, but several approaches have been found to work in specific situations (e.g., [65][66][67]). A differential sensor scheme can be used to (imperfectly) subtract out the nonspecific component of the sensor response (see Sec.…”

Section: Selectivitymentioning

confidence: 99%

Label‐Free Impedance Biosensors: Opportunities and Challenges

2007

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Impedance biosensors are a class of electrical biosensors that show promise for point-of-care and other applications due to low cost, ease of miniaturization, and label-free operation. Unlabeled DNA and protein targets can be detected by monitoring changes in surface impedance when a target molecule binds to an immobilized probe. The affinity capture step leads to challenges shared by all label-free affinity biosensors; these challenges are discussed along with others unique to impedance readout. Various possible mechanisms for impedance change upon target binding are discussed. We critically summarize accomplishments of past label-free impedance biosensors and identify areas for future research.

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Section: Selectivitymentioning

confidence: 99%

Label‐Free Impedance Biosensors: Opportunities and Challenges

2007

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“…We recently reported the specificity of each monoclonal antibody when tested with a panel of 8 different gangliosides and a standardized ELISA protocol. 19,20 Gangliosides were separated on aluminum-backed silica gel plates, and the plates were dipped in 0.2% polyisobutyl-methacrylate in hexane for 1 min. After drying, the plates were blocked with PBS-1% HSA for 30 min, washed with PBS and dried.…”

Section: Immunostaining Of Thin-layer Chromatogramsmentioning

confidence: 99%

“…20 This ELISA considers the following factors: (i) microtiter plate type; (ii) purity of the commercial gan-glioside; (iii) antigen-coating strategy; (iv) nature of protein in the blocking buffer; (v) concentration and nature of the detergent in the wash buffer; (vi) background; and (vii) positive and negative controls. 23 Briefly, the microtiter plates (Falcon Probind 3915) were coated with an ethanolic suspension of gangliosides (3 nmol/ 100 lL/well) (Sigma) and dried in a vacuum desiccator for 2 days.…”

Section: Quantitation Of Cell-surface Antigensmentioning

confidence: 99%

Endogenous immune response to gangliosides in patients with confined prostate cancer

Ravindranath

Muthugounder

Presser

et al. 2005

Intl Journal of Cancer

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Our study investigated whether endogenous IgM antibodies to gangliosides occur in patients with early stages of prostate cancer (CaP) patients, after defining ganglioside profiles of CaP cell lines. Immune and resorcinol staining detected the presence of gangliosides GM3, GM2, GD3, GD2 and GD1a but not GM1a, GD1b or GT1b in the extracts of normal prostatic epithelial cells (PrEC) and neoplastic androgen‐insensitive (PC‐3, DU145) and ‐sensitive (LNCaP‐FGC and LNCaP‐FGC‐10) CaP cells. Using a sensitive ELISA, developed and validated in our laboratory, the titers of IgM against 8 gangliosides from sera of patients with benign prostatic hyperplasia (BPH) (n = 11), organ‐confined (T1/T2, n = 36) and unconfined (T3/T4, n = 27) CaP and age‐matched healthy men (n = 11) were determined double‐blinded. Using ANOVA and Fisher's least significant difference (LSD) methods, the log‐titers among different groups were compared. CaP patients differed from healthy and BPH patients in increased titers against GD1a and decreased titers against GD3. Titers of antibodies to other gangliosides exhibited no difference between CaP patients and others. The specific augmentation of anti‐GD1a IgM in patients with organ‐confined CaP (stage T1/T2) but not in patients with unconfined CaP (stage T3/T4) or BPH or in healthy controls is striking. This finding together with identification of GD1a as a major ganglioside in CaP cell lines and with the accruing studies on the immunosuppressive nature of GD1a indicates that augmentation of anti‐GD1a IgM in confined CaP may signify an early endogenous immune response to eliminate a “danger signal” from tumor microenvironment and circulation. © 2005 Wiley‐Liss, Inc.

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“…Many studies have been already done to improve the ELISA itself [6][7][8][9], so that the methodology of indirect ELISAs is well established. However, there is as yet no generally accepted way of expressing the ELISA results from such studies.…”

Section: Introductionmentioning

confidence: 99%

Development and characterization of a standardized ELISA including a reference serum on each plate to detect antibodies induced by experimental malaria vaccines

Miura

Orcutt

Muratova

et al. 2008

Vaccine

201

188

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Enzyme linked immunosorbent assay (ELISA) has been widely used to measure antibody titers for evaluating the immunogenicity of a vaccine. However, there is as yet no generally accepted way of expressing the ELISA results in the case of experimental vaccines, since there is usually no uniform standard. Both end point and single dilution methods have significant disadvantages. In this paper, we obtained reproducible data with fewer dilutions of samples by addition of serially diluted standard serum to each ELISA plate. Since this ELISA method gives reliable antibody titer with less labor than other methods, it can strongly support vaccine development.

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Factors affecting the fine specificity and sensitivity of serum antiganglioside antibodies in ELISA

Cited by 52 publications

References 43 publications

Label‐Free Impedance Biosensors: Opportunities and Challenges

Label‐Free Impedance Biosensors: Opportunities and Challenges

Endogenous immune response to gangliosides in patients with confined prostate cancer

Development and characterization of a standardized ELISA including a reference serum on each plate to detect antibodies induced by experimental malaria vaccines

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