Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1995-09-29
2000-10-31
Scheiner, Laurie
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 691, 435 693, 435 701, 530350, 4241881, 4242081, G01N 3353
Patent
active
061400599
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention is relative to a method for the obtention of native domains of viral membrane proteins, especially of native ectodomains of the surface protein gp 160 of the human immunodeficiency virus HIV, the causative agent of AIDS (acquired immune deficiency syndrome), as well as the native protein domains themselves obtained by this method and their use as vaccine, especially the use of native ectodomains of the surface protein gp 160 of the human immunodeficiency virus HIV as vaccine against HIV. A method for determining suitable sites positions for the insertion of recognition sequences into the protein domains to be obtained, which function as enzymatic cleavage sites, especially as collagen splitting sites, as well as recognition sequences for collagenases themselves, is also a component of this method for the obtention of native domains of viral membrane proteins.
BACKGROUND OF THE INVENTION
A proven means for the prophylaxis of viral diseases is immunization, the vaccination of the body against the viruses. In earlier times viral diseases which occurred in epidemic fashion such as e.g. smallpox and polio were frequently able to be checked by means of vaccination. In order to achieve such an active immunization against a virus the body, more precisely the immune system, is exposed to a viral antigen. This can take place e.g. by injecting inactivated or attenuated viruses or also parts, e.g. of proteins of the virus. The human immune system is basically composed of two different partial systems, the cellular immunological response (T cells) and the humoral immunological response (B cells, antibodies). In the case of a reinfection with a virus the humoral immunological response, imparted by B cells, represents the most important defense mechanism.
The immune system of the body reacts to the antigen with the formation of specific antibodies which recognize and bind the antigen and therewith the virus, thus initiating its inactivation. In addition, so-called "memory cells" are formed, that is, special lymphocytes which are activated upon a later infection with "their" virus or "their antigen" and stimulate the immune system very rapidly to synthesize large amounts of the antigen-specific antibodies. In this manner the immune system can react significantly more rapidly to a viral attack than if it had never been confronted previously by the corresponding antigen.
However, the use of attenuated viruses for vaccination can be problematic in as far as it can not be excluded that the viruses used become virulent in the body again and an outbreak of the viral disease occurs. It is preferable for this reason to use an isolated protein of the virus as immunogen. For this, it is best to use a surface protein of the virus since it is normally readily accessible to the humoral immunological response. Ideally, only a part of a surface protein is used, namely, that part which is located externally on the virus surface, the so-called ectodomain, since in the case of the intact virus only this part is accessible to antibodies. However, care should be taken in the production of a protein domain to be used for vaccination against a virus that it should have the native form to the extent possible since otherwise there is the danger that antibodies are formed which do not recognize the native protein and therewith the virus and are therewith ineffective for neutralization. Such molecules or molecule parts which are recognized on the basis of their three-dimensional structure by antibodies are called conformational or structural epitopes and those which are recognized by their amino-acid sequence are called sequential epitopes. It turns out more and more that there are only very few sequential epitopes and that most antigenic epitopes are structural, that is, they must exhibit the correct three-dimensional folding of the polypeptide chain in order to be recognized by corresponding antibodies. "Native" means in this context that the spacial structure of the protein domain used is identic
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Parkin Jeffrey S.
Scheiner Laurie
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