Colloid compositions for solid phase biomolecular analytical, preparative and identification systems
a biomolecular analytical and solid-phase technology, applied in the field of colloidal compositions for solid-phase biomolecular analytical, preparative and identification systems, can solve the problems of laborious and labor-intensive preparation of the prior art microarray system, and achieve the effect of convenient and convenient us
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example i
Preparation of a Generic Composition According to the Invention and Use of the Composition for Preparation of a Microarray
[0033] The first step in implementing the improvements possible with assay systems according to the invention is to prepare a colloidal suspension of defined particle size matrix material from, e.g., a highly charged material such as nitrocellulose, activated nylon or polyvinylidene difluoride. These materials are capable of binding or adsorbing a variety of charged molecules such as nucleic acids, proteins, etc., and such binding is sufficiently strong to survive the high stringency conditions required for hybridization reactions. The matrix material in colloidal suspension could also be a non-charged polymer, such as plastic, nylon or polysulfone. The colloids are prepared by using any number of traditional methods that may include pulverization, precipitation and spray drying, as well as a variety of other well-established techniques.
[0034] To aliquots of th...
example ii
Determination of Protein / Ligand Interactions
[0046] Binding reactions between proteins and ligands can be evaluated easily using the system of the invention, e.g., as a means of obtaining useful information for diagnostic or research purposes. Furthermore, in drug discovery and development, the measured ligand / protein interactions can be determined in the presence of potential inhibitors or enhancers of selected protein / ligand binding. Specifically, e.g., antigen, for example, hepatitis B surface antigen, tetanus toxoid or viral antigens can be bound to the colloid matrix particles and used to detect specific antibody. Also, specific antibody can be bound to colloid to detect antigen.
[0047] In one embodiment, a spotting composition is preparedaccording to Example I to include a protein of interest or a segment thereof, that contains a ligand binding site. The composition is then treated with non-specific blocking agents, as in Example I. Microarray spotted slides are formed by depo...
example iii
HLA Typing
[0051] Although the membrane based system of the prior art has been a useful research tool and has been used for routine human leukocyte antigen (HLA) typing analysis, it is a cumbersome, manual procedure that is labor intensive and inefficient. Using the system of the invention, however, a much more convenient method for routine HLA typing can be devised. The new method comprises conventional isolation of genomic DNA from an individual and amplification of the HLA gene of interest using specific HLA primers. In addition, an HLA microarray can be made with each spot being formed from a liquid colloidal composition according to this invention, wherein the probe of each spot is a sequence specific oligonucleotide that is complementary to a region of a known HLA polynucleotide sequence. A plurality of such probes representing a plurality of the hypervariable regions were used. For example, for intermediate level resolution HLA-DRB1 typing, 32 probes, characterized in a “Work...
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Abstract
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