HUMAN LOW FC EPSILON-RECEPTOR,THE ISOLATION,THE RECOMBINANT PREPARATION AND PURIFICATION THEREOF

• Main Authors: HITOSHI KIKUTANI, EDWARD L. BARSUMIAN, TADAMITSU KISHIMOTO, MASAKI SUEMURA
• Format: Patent
• Language: English
• Published: 26.04.1989
• Edition: 4
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; HUMAN NECESSITIES; HYGIENE; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; MEDICAL OR VETERINARY SCIENCE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The present invention is concernd with a) isolation and purification of the soluble IgE binding factor (Fc epsilon R) secreted or shed by lymphoblastoid cells, b) partial sequencing the obtained Fc epsilon R-receptor, isolating the thus obtained fragments and determination of the sequences of the thus obtained fragments, c) preparation of an oligonucleotide encoding to one of the said partial sequences, d) isolating and identifying expression vehicles containing the gene coding for Fc epsilon -receptor, comprising the steps of: synthesizing cDNA from a RNA matrix derived from lymphoblastoid cells producing Fc epsilon -receptor mRNA, incorporating said synthesized cDNA in expression vehicles to form an expression vehicle bank, hydribizing said incorporated cDNA to identify those expression vehicles which contain a gene coding for Fc epsilon -receptor, with a labeled probe comprising an oligonucleotide common to the gene for Fc epsilon --receptor, replication and expression of the thus obtained gene, and sequencing the thus obtained gene, e) determination of the gene coding for the active human low-affinity Fc epsilon -receptor utilising isolated cDNA sequence obtained from the vehicles from operation d), f) expression of the Fc epsilon R cDNA in Cos 7 cells and g) preparing an expression vector containing the DNA sequences coding for a soluble fragment of the Fc epsilon -receptor and expressing said soluble fragments in a microorganism or in mammalian cells. The water-soluble fragment of the Fc epsilon -receptor is suitable for treatment of allergic reactions.