Abstract
The article presents the results of research to obtain specific components of the preparation selective biosensor surface for further development the immune biosensor test-system for the express – indication of bacteria Pseudomonas aeruginosa and their detection in biological material and in the environment. Specific antiserum and immunoglobulins to P. aeruginosa were obtained by autors. The resulting surface modification components for activity and specificity of the specific compounents were tested using analytical device imunosensor «Plasmon-6», based on surface plasmon resonance. Biosensors are defined as any measuring device that contains a biological element. It combines the exquisite selectivity of biology with the processing power of modern microelectronics and optoelectronics to offer powerful new analytical tools with major applications in the field of medicine, environmental studies, food and processing industries. These analytical devices are based on the union between biological and physio-chemical components. Biological components include macro-molecules such as antibodies, enzymes, tissue slices which are used to recognize and interact with a specific analyte. Physiochemical components are usually referred to as transducers which converts the interactions into signals; it is later amplified with respect to its concentration of analyte. The transducer may use potentiometric, amperometric, optical, magnetic, colorimetric devices. A target analyte in the external membrane must be able to enter the biosensor. The external membrane of the biosensor must be permeable to the analyte where the biosensor is sensitive to it. The biological element inside the biosensor then interacts with chemical species through a biochemical reaction which in turn produces another chemical product and characterized by change in mechanical, electrical properties. The output signal may be a conventional electrochemical signal depending on the type of transducer it uses. Assessment of P. aeruginosa was carried out using an analytical device - immunosensor, with immobilized specific antibodies on the transducer surface. The antibodies have interact with cell antigens, and the resulting shift value resonance angle recorded. Changing the angle depends on the amount of the immune complexes formed on the transducer surface. From the obtained results on the selective surface of transducer of the imunosensor, we can see, that the diagnostic system works with IgG concentration of 1 mg/ml, working titer of 1:7 in polyclonal antibodies against P. aeruginosa. The resulting antiserum specific immunoglobulins can be used in preparing the selective surface of immune biosensor.
Highlights
Scientific Messenger of Lviv National University of Veterinary Medicine and Biotechnologies named after S.Z
Отримані компоненти модифікації поверхні було протестовано на активність та специфічність за допомогою аналітичного приладу імуносенсора на основі поверхневого плазмонного резонансу «Плазмон-6»
Specific antiserum and immunoglobulins to P. aeruginosa were obtained by autors
Summary
Кролів імунізували за схемою грундімунізації, що ґрунтується на введенні 1 см антигену (штам P. aeruginosa ATCC 9027) з повним ад’ювантом Фрейнда (1:1) уздовж хребта у 6 точок підшкірно з інтервалом два тижні. Через 45 діб проводили гіперімунізацію за схемою: введення підшкірно антигену спочатку з неповним адьювантом, а потім без ад’юванту в дозі 0,1 см з десятиденним інтервалом, впродовж трьох тижнів. До введення антигену у тварин-донорів не було витікають з колонки, визначали з допомогою 5 %-го виявлено специфічних антитіл до P. aeruginosa, але розчину трихлороцтової кислоти. Наявність солей уже після двократного введення полівалентного антиамонію у фракціях, що витікають, визначали якісною гену виявлено стрімке утворення антитіл (табл.). Імунізованих за вищезазначеною схемою, визначали на 55, 65 та 75-у добу. Динаміка титрів антитіл у кролів під час гіперімунізації їх антигеном P. aeruginosa в реакції аглютинації
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