Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Patent
1994-04-05
1998-09-15
Eisenschenk, Frank C.
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
4353201, 435 91, 5303873, 5303883, 5303871, 5303894, C12N 1513, C07H 2104
Patent
active
058080325
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to anti-hepatitis B virus surface antigen (hereunder abbreviated as "HBs") antibody genes and expression plasmids therefor. More particularly, this invention relates to polynucleotides that respectively code for the L and H chains of a human anti-HBs antibody immunoglobulin and portions thereof, as well as expression plasmids containing those polynucleotides.
BACKGROUND ART
Hepatitis B virus is a dreadful source of infection which not only causes acute hepatitis by infection but also aggravates it, by continued infection, to irrecoverable chronic hepatitis, sometimes to cirrhosis and even to liver cancer. Three major types are known as antibodies against hepatitis B virus and they include an anti-HBs antibody, as well as an antibody against the core antigen (HBc) within the virus particle and an antibody against the e antigen (HBe) which is held to be contained in the core antigen. Among these three antibodies, the anti-HBs antibody has the hepatitis B virus neutralizing activity and, hence, it is expected to exhibit the phylactic effect against the virus and the ability to prevent induced manifestation of the disease and there exists a very strong demand that pharmaceutical preparations of the antibody be marketed as soon as possible.
However, producing pharmaceutical preparations of the anti-HBs antibody using human HBs antibody positive blood plasma as a starting material is subject to constraints on the starting material and, hence, is not practical. Monoclonal antibodies can be produced by using the cell fusion technique which has seen rapid developments in recent years but there are some doubts about the safety of the mouse derived antibody in administration to the human body. Furthermore, it is difficult to prepare a proliferative hybridoma that is stable for a sufficiently long time to achieve large-scale production of the human derived monoclonal antibody. Thus, none of the conventional techniques are capable of consistent supply of the human anti-HBs antibody as a pharmaceutical preparation.
DISCLOSURE OF INVENTION
The object of the present invention is to provide a technique by which a pharmaceutical preparation of the anti-HBs antibody that may safely be administered to the human body can be produced easily in a large amount and without any problems associated with the procurement of starting materials.
Thinking that the production of a human immunoglobulin with high titer and selectivity in microorganisms would be useful for the development of a pharmaceutical preparation of immunoglobulin for use in passive immunotherapy, the present inventors repeated intensive studies with a view to accomplishing large-scale production of the human monoclonal anti-HBs antibody in microorganisms. As a result, they succeeded in cloning human monoclonal anti-HBs antibody genes and expressing said genes by the recombination DNA technique. The present invention has been accomplished on the basis of this success.
Stated more specifically, the present invention relates to polynucleotides that code for the L and H chains of the human anti-HBs antibody immunoglobulin, as well as polynucleotides coding code for polypeptides that comprise portions of said L and H chains, respectively, which when expressed are reasonably effective in passive immunity (e.g., phylaxis against HB virus, prevention of manifestation of the disease and its treatment). The present invention also relates to plasmids that are capable expressing the L and H chain genes of the human anti-HBs antibody immunoglobulin and polypeptides that are reasonably effective in passive immunity, as well as expression plasmids containing said polynucleotides.
Screening for polypeptides that are reasonably effective in passive immunity can be readily accomplished by using a known assay technique such as enzyme immunoassay with the HBs antigen or passive hamagglutination test (a test kit such as "Hebsgencell" is commercially available from The Green Cross Corporation), with the anti-HBs titer being detected either qua
REFERENCES:
GeneBank report pp. 14, 15, 29 and 30.
Arima Kenji
Kurihara Tatsuya
Matsukura Shigekazu
Nishihara Tatsuro
Tsuruoka Nobuo
Eisenschenk Frank C.
Suntory Limited
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