Abstract: Provided are a detection method for a myriad of proteins involved in an autoimmune disease with high sensitivity and high efficiency, and an analysis method for data resulting from the detection method. In order to construct the detection method and analysis method, there is provided means for comprehensively analyzing the proteins involved in an autoimmune disease by bringing a mammal-derived protein expressed in a cell-free protein synthesis system into contact with a sample derived from a patient with an autoimmune disease to detect autoantibody production, and subjecting the detected data to statistical analysis processing, and further, gene ontology analysis and/or pathway analysis.

Abstract: The present invention provides a method for preparing cell extract for high-performance cell-free protein synthesis reactions, comprising the elimination of low molecular weight substances that have protein synthesis inhibitory activity, from cell extracts that have cell-free protein synthesis activity, by such methods as dialysis, gel filtration and ultrafiltration. Also provided is a ready-made cell-extract for cell-free protein synthesis employing this method. Furthermore, the formation of insoluble matter can be reduced by performing the process for eliminating the low molecular weight substances that inhibit protein synthesis in the co-presence of ATP, GTP or amino acids, so as to stabilize the cell extract.

Abstract: Provided are a detection method for a myriad of proteins involved in an autoimmune disease with high sensitivity and high efficiency, and an analysis method for data resulting from the detection method. In order to construct the detection method and analysis method, there is provided means for comprehensively analyzing the proteins involved in an autoimmune disease by bringing a mammal-derived protein expressed in a cell-free protein synthesis system into contact with a sample derived from a patient with an autoimmune disease to detect autoantibody production, and subjecting the detected data to statistical analysis processing, and further, gene ontology analysis and/or pathway analysis.

Abstract: The present invention relates to a malaria vaccine comprising: (a) a polypeptide consisting of an amino acid sequence of SEQ ID NO: 1, 2, or 3; (b) a polypeptide consisting of an amino acid sequence of SEQ ID NO: 1, 2, or 3, wherein one or more amino acids are deleted, substituted and/or added and having effect for preventing falciparum malaria; or (c) a polypeptide consisting of an amino acid sequence having 70% or more identity with an amino acid sequence of SEQ ID NO: 1, 2, or 3 and having effect for preventing falciparum malaria.

Abstract: Methods to construct a transcription template for cell-free protein synthesis that has a high translation template activity, using a 3?-side primer and a 5?-side primer for PCR are provided. The 5?-side primer for PCR has a sequence complementary to a base sequence containing at least a part of a promoter functional site from the 5?-end of a promoter and has a base sequence that does not contain a base sequence complementary to at least a part of a RNA polymerase-recognizing site of the 3?-end of the promoter. The other primer has a base sequence complementary to at least a part of the RNA polymerase-recognizing site of the 3?-end of the promoter and has a sequence that does not contain a base sequence complementary to at least a part of a promoter functional site from the 5?-end of the promoter.

Abstract: The present invention presents construction of a detection method requiring no step of removing free biotin during preparation of a biotinylated protein having a biotin tag, in a detection method of a substance interacting with a protein, and studied various preparation methods of the biotinylated protein. In order to solve the above-mentioned problem, the present inventor has found that in a cell-free protein synthesizing system, in particular, a wheat embryo cell-free protein synthesizing system, when biotinylation is performed during or after protein's synthesis, the biotinylation of the protein can be attained in an remarkably lower concentration of the biotin than that in the conventional biotinylation operations, and has accomplished the present invention by using the protein having the biotin tag in each detection system.

Abstract: The present invention is intended to provide a safe and quick means for screening a drug to a bioactive protein, in particular, an inhibitor, using a cell-free protein synthesis system with the use of a wheat embryo extract solution. The present inventors have strenuously studied to solve the matters above and finally completed the present invention by, in a system with the use of a wheat embryo, among cell-free protein synthesis means, constructing a synthesis system of a bioactive protein while sustaining its activities, and constructing a system for screening an inhibitor candidate to SARS 3CLpro, as an example using the synthesis system.

Abstract: The present invention provides a cell-free protein synthesis method characterized by a cell-free protein synthesis reaction solution that is weakly reducing and suitable for protein folding, performing a weakly reducing synthesis reaction, and preferably performing a translation reaction with the further addition of a substance catalyzing a disulfide bond exchange reaction at the beginning of the translation reaction. This method allows for proper formation of intramolecular disulfide bonds in the protein and efficiently provides protein having substantially the same function as the original protein. Specifically, it provides antibody protein that binds specifically to an antigen.

Abstract: A method of detecting a reaction between a fluorescently labeled protein synthesized in a cell-free protein synthesizing system and a sample solution simply, in a short time and at high precision is provided. A case of detecting a binding reaction between an antibody fused with GFP and a sugar on the nanoparticle surface is explained. In a well of a microplate, a solution containing an antibody fused with GFP, and a solution containing a nanoparticle with a sugar reactive with the antibody fused with GFP adhered to a surface thereof are mixed to prepare a mixed solution A, and after the reaction, FCS measurement is performed.

Abstract: The present invention presents construction of a detection method requiring no step of removing free biotin during preparation of a biotinylated protein having a biotin tag, in a detection method of a substance interacting with a protein, and studied various preparation methods of the biotinylated protein. In order to solve the above-mentioned problem, the present inventor has found that in a cell-free protein synthesizing system, in particular, a wheat embryo cell-free protein synthesizing system, when biotinylation is performed during or after protein's synthesis, the biotinylation of the protein can be attained in an remarkably lower concentration of the biotin than that in the conventional biotinylation operations, and has accomplished the present invention by using the protein having the biotin tag in each detection system.

Abstract: Disclosed are a preparation containing cell extracts for cell-free protein synthesis, prepared by excluding from a living organism a system, participating to inhibiting of self protein synthesis reaction, an apparatus for cell-free protein synthesis reaction equipped with a reaction tank for cell-free protein synthesis, and a kit for use therefor; the preparation can be stored at room temperature and prepared as a preparation in a state where biological functions of the cell extracts are maintained, and further, disclosed is methods for cell-free protein synthesis comprising providing cell extracts from which an inhibitor for self protein synthesis reaction is substantially excluded, having introduced therein treatment selected from supplement, storage, exchange or discharge with respect to an element selected from at least mRNA serving as a template for synthesis reaction, an energy reproduction system enzyme, a substrate, and an energy source.

Abstract: The subjects of the present invention are to prepare a highly functionalized cell extract for cell-free protein synthesis and to specify and eliminate inhibitors and unstable substances present in conventional and various cell extracts for cell-free protein synthesis. Also provided is a method for preparing the cell extract for use in a cell-free protein synthesis means, wherein ATP-mediated phosphorylation pathway of sugar present in the cell extract is controlled. In particular, the control is introducing at least one selected from the following: 1) removing monosaccharides, 2) removing phosphorylated sugars, 3) controlling production of monosaccharides from polysaccharides, and 4) controlling production of phosphorylated sugars from monosaccharides.

Abstract: An object of the present invention is to provide a novel means for finding a passive substance to the action of a selected trigger protein in a target cell system. In particular, the present invention succeeded in establishing a novel method for screening an indicator substance by the steps of contacting the trigger protein with a target cell extract to initiate the action on an unspecified indicator substance by the trigger protein, and specifying the substance changed by the action induced by the trigger protein. The primary part of the present invention comprises the following steps: 1) contacting a trigger protein prepared by a cell-free protein synthesis means with a target cell extract which contains the indicator substance that is passively produced by an action induced by the trigger protein and desired to screen, to initiate the action by the trigger protein, and 2) specifying the substance changed by the action induced by the trigger protein.

Abstract: To develop system technology for a high throughput in vitro synthesis reaction method for biopolymers such as proteins, RNA, and the like.

Abstract: An object of the present invention is to provide a means for producing an antigenic component with the retained native antigenicity using a cell-free protein synthesis. In particular, it is an object to provide a means for producing an antigenic component without depending on codon usage, like expressing an antigenic component from a gene containing a large amount of AT. The present inventors have made a strenuous study to solve the matters described above and successfully completed the present invention by preparing an antigenic component with the retained antigenicity, in particular a malaria antigen useful for manufacturing a malaria vaccine, through a system with the use of a wheat embryo among cell-free protein synthesis means.

Abstract: An object of the present invention is to provide a reagent for protein synthesis by a simple operation which is advantageous in a cell-free protein synthesis in many samples in a test using an aimed (specific) protein or a high throughput analysis using a protein chip, etc. and also to provide a method for the synthesis of protein using the same. Further objects are to provide a protein library using the above-mentioned reagent, a kit for preparing a protein chip, a kit for testing an aimed (specific) protein, etc.

Abstract: The present invention is intended to provide a safe and quick means for screening a drug to a bioactive protein, in particular, an inhibitor, using a cell-free protein synthesis system with the use of a wheat embryo extract solution. The present inventors have strenuously studied to solve the matters above and finally completed the present invention by, in a system with the use of a wheat embryo, among cell-free protein synthesis means, constructing a synthesis system of a bioactive protein while sustaining its activities, and constructing a system for screening an inhibitor candidate to SARS 3CLpro, as an example using the synthesis system.

Abstract: The present invention provides a single chain antibody that retains its original specific binding activity with an antigen, and a labeled single chain antibody in which a labeling substance is bound to the single chain antibody. Specifically, the labeled single chain antibody of the present invention can be produced by linking a labeling substance to a linker part of a single chain antibody. The antibody is produced using a wheat embryo-derived cell-free protein synthesis system, and production is carried out in a low reductive state that allows an intramolecular disulfide bond to be retained. Further, bonding the antibody to a solid phase via the labeling substance enables production of an immobilized single chain antibody as well as a method for analyzing an antigen-antibody reaction using the immobilized single chain antibody.

Abstract: Utilizing a what embryo cell-free protein synthesis system, there are provided a process for the production of selenomethionine-labeled protein, characterized in that, methionine in a wheat embryo extract for a cell-free protein synthesis obtained by a complete removal of endosperm contaminated is changed to selenomethionine and a cell-free protein synthesis is carried out using a reaction solution composition for protein synthesis containing selenomethionine instead of methionine under a batch condition or a dialysis condition and also the said protein produced as such. There are further provided a process for the production of heavy hydrogen-labeled protein using the same means and also the said protein produced as such.

Abstract: The present invention provides a composition for cell-free protein synthesis, which is superior in storage stability in a freeze-dried state, more particularly a freeze-dryable or freeze-dried composition for cell-free protein synthesis, which contains a cell extract for cell-free protein synthesis and inositol, and a freeze-dryable or freeze-dried composition for cell-free protein synthesis containing a cell extract for cell-free protein synthesis, and a deliquescent material in a proportion of not more than 0.01 part by weight per part by weight of a protein in the composition; and a composition for cell-free protein synthesis superior in storage stability in a frozen state, more particularly a freezable or frozen composition for cell-free protein synthesis, containing a cell extract for cell-free protein synthesis and polyhydric alcohol.