A homogeneous immunoassay for the mycotoxin T-2 utilizing liposomes, monoclonal antibodies, and complement

Ligler, Frances S.; Bredehorst, Reinhard; Talebian, Abdolhossen; Shriver, Lisa C.; Hammer, Charles F.; Sheridan, James P.; Vogel, Carl‐Wilhelm; Gaber, Bruce P.

doi:10.1016/0003-2697(87)90237-5

Cited by 29 publications

(10 citation statements)

References 13 publications

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“…Тому Т-2 токсин необхідно було модифікувати, тобто ввести в його структуру реакційноздатну групу, за допомогою якої Т-2 токсин міг би ковалентно з'єднатися з протеїновою молекулою. Згідно з даними літератури [6,12], для одержання модифікованого Т-2 токсину (гаптену) його піддають взаємодії з бурштиновим ангідридом. При цьому ангідрид бурштинової кислоти реагує з гідроксильною групою Т-2 токсину з утворенням складного ефіру, а друга карбоксильна група залишається вільною і може бути використана для приєднання модифікованого Т-2 токсину до молекули протеїну (рис.…”

Section: модифікація т-2 токсину бурштиновим ангідридомunclassified

“…Вихідний Т-2 токсин має температуру плавлення 147-147,5 °С. На жаль, в літературі [6,12] немає даних щодо агрегатного стану активованого Т-2 токсину. Для відпрацювання оптимальних умов його синтезу повторили синтез без нагрівання реагентів, аналогічно роботі [12].…”

Section: модифікація т-2 токсину бурштиновим ангідридомunclassified

“…Надалі для визначення титру антитіл застосовували більш чутливі методи. Методом конкурентного імуноензимного аналізу (ELISA) було встановлено [12], що сироватки крові, отримані при імунізації кролів кон'югатом Т-2 токсину з бичачим сироватковим альбуміном з титром антитіл 1:9600 дозволяють досягти межі виявлення Т-2 на рівні 10 нг/мл. Імуносенсорний аналіз дозволяє визначати Т-2 токсин в розчинах у лінійному діапазоні концентрацій від 1 нг/мл до 1000 нг/мл.…”

Section: отримання поліклональних антитіл до т-2 токсинуunclassified

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Synthesizing t-2 toxin conjugates with proteins for the use in immunosensitive test systems

Гойстер

Кривенчук²,

Хмельницький

2015

EEJET

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Section: модифікація т-2 токсину бурштиновим ангідридомunclassified

Section: отримання поліклональних антитіл до т-2 токсинуunclassified

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Synthesizing t-2 toxin conjugates with proteins for the use in immunosensitive test systems

Гойстер

Кривенчук²,

Хмельницький

2015

EEJET

View full text Add to dashboard Cite

“…or other pathogen strains of microorganisms through nucleic acid hybridization. So far detection of Mycobacterium has been reported with dye-encapsulating liposomes bearing mycotoxin T2, 30 after lysis of the lipid bilayer by complement activation, with liposomes bearing mycolic acid for the detection of the specific antigens 31 format. 32 The concept was to prepare and characterize liposomes consisting of EggPC (Egg phosphatidylcholine) and DPPG (1,2-Dipalmitoyl-sn-Glycero-3-[Phospho-rac-(1-glycerol)] (Sodium Salt)) with a simple to synthesize thiol-reactive conjugate (Mal-SA) incorporated in their bilayer.…”

Section: Introductionmentioning

confidence: 99%

Development and Characterization of Oligonucleotide-Tagged Dye-Encapsulating EPC/DPPG Liposomes

Mourelatou

Spyratou

Georgopoulos³

et al. 2010

J. Nanosci. Nanotech.

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Liposomes applications in health care include meanly their ability to carry drugs and genes inside the human body for therapeutic purposes. Nevertheless their applicability can extend far beyond and could be used as analytical tools in order to perform rapid, low-cost, sensitive and specific analyses. Their physical characteristics, such as large internal volume and extended surface area, render them ideal for these applications and specifically for improving the specificity and sensitivity of the analytical assay. The purpose of this study was to develop a simple, stable and low-cost oligonucleotide-tagged liposomal formulation consisting of EggPC and DPPG with a simple to synthesize thiol-reactive conjugate (Mal-SA) incorporated into the lipid bilayer of liposomes. The prepared liposomes, having also the water soluble dye Sulforhodamine B encapsulated in their inner cavity, were characterized in terms of their physicochemical (size, size distribution, zeta-potential, lipid content) and mechanical (morphology, rigidity) properties. The results showed that the final liposomal formulation could be used in the future as analytical tool for detecting pathogen strains of microorganism in biological milieu.

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“…Antibody or antigen can be attached to the liposome's surface, while each liposome can entrap relatively large amounts of a measurable substance in the aqueous interior. The entrapped material can be a fluorophor (1)(2)(3)(4)(5) or enzyme (6-12). The liposomes and their contents can be made to act as the nmarkerslt in the immunoassay as an alternative to other labels such as isotopes or enzymes directly coupled to antibodies.…”

Section: Introductionmentioning

confidence: 99%

A Novel Colourimetric Homogeneous Liposomal Immunoassay Using Sulphorhodamine B

Frost

Firth

Chakraborty

1994

Journal of Liposome Research

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The dye, Sulphorhodamine B, was entrapped within liposomes prepared by the reverse evaporation technique. Marked differences in absorption spectra were found when free and entrapped dyes were compared, with a shift in the wavelength of maximum absorption. When entrapped dye was released by lysis of the liposomes, for example by detergent, the absorption spectrum reverted to that of free dye. This absorption change was employed in a novel marker system for complement-mediated immunoassay. As a model assay, human serum albumin was measured using this approach. Liposomes which had been coated with albumin were incubated with anti-albumin antibody and complement and the resulting absorption change measured using an automated spectrophotometric analyser. The decrease in absorption change on the addition of albumin formed the basis of a competitive homogeneous immunoassay for human serum albumin. Using purified albumin as a standard, a correlation of 0.96 was obtained when the albumin in human serum was measured in the liposomal assay and the results compared to measurements using a bromocresol green method.

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A homogeneous immunoassay for the mycotoxin T-2 utilizing liposomes, monoclonal antibodies, and complement

Cited by 29 publications

References 13 publications

Synthesizing t-2 toxin conjugates with proteins for the use in immunosensitive test systems

Synthesizing t-2 toxin conjugates with proteins for the use in immunosensitive test systems

Development and Characterization of Oligonucleotide-Tagged Dye-Encapsulating EPC/DPPG Liposomes

A Novel Colourimetric Homogeneous Liposomal Immunoassay Using Sulphorhodamine B

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