Process for disintegrating nucleic acids and preparing biologica

Details Process for disintegrating nucleic acids and preparing biologica Process for disintegrating nucleic acids and preparing biologica

1998-11-18

2000-12-26

Park, Hankyel

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving virus or bacteriophage

435 6, 435325, 424 931, C12Q 170, C12Q 168, C12N 500, A01N 6300

Patent

active

061657115

DESCRIPTION:

BRIEF SUMMARY
BACKGROUND OF THE INVENTION

1. Field of the Invention
The present invention relates to the disintegration of biologically active nucleic acids in a biologically active material by exposing the biological material, optionally treated with a photodynamic substance, to a laser beam which yields a non-infectious biological product having essentially all biologically active nucleic acid disintegrated while the biological integrity and activity, such as antigenicity, immunogenicity, protectivity or enzymatic characteristics, of the biological material is maintained. The invention relates also to the production of biologicals, such as vaccines, genetically engineered protein products, monoclonal antibodies or blood factors having essentially all biologically active nucleic acid disintegrated.
The use of continuous cell lines (CCL) for the production of therapeutic biological products and the problems of contaminating nucleic acids in biological products as well as the inactivation of residual contaminating cellular DNA in such biologicals have been discussed. While certain products, such as recombinant proteins, vaccines or monoclonal antibodies, are effectively produced using vertebrate cells, certain other protein products are not easily obtained from common used cells due to the low production rate of the proteins in those cells. Other mammalian cells which are easy to cultivate are not licensed by regulatory authorities for the production of genetically engineered proteins and for their use for therapeutic and prophylactic porposes in humans or animals.
It has been suggested to use mammalian cell lines for the genetically engineered manufacture of many proteinaceous products as well as for the production of virus vaccines. However, certain problems are anticipated in the use of such cell lines. Many products are not released or secreted from mammalian cells directly into the culture medium. Accordingly, the harvest of such products often will require rupturing cellular membranes to release those products into the medium from which they may subsequently be refined or purified. Such rupture, however, will also release cellular nucleic acid into the medium. Particularly because many easily cultured cell lines are transformed mammalian tumorogenic cell lines with an oncogenic potential, such as MDCK cells or SK-Hep cells, a need exists to ensure that active nucleic acid is not present as a contaminant in the proteinaceous end product or in the vaccine preparation. Attempts, therefore, have been made to inactivate cellular nucleic acids that might be associated with a risk of oncogenicity.
Vaccination against viral diseases has been one of the major tasks of medicine over the past century. While effective vaccines have been developed for a large number of diseases, development of safe and effective vaccines for a number of other diseases, e.g. HIV infections, remain problematic. The use of inactivated or killed viral agents as a vaccine, although generally safe, will not always be effective if the immunogenic characteristics of the agent are altered. However, if a highly infectious, virulent virus is used for vaccine manufacture, one has to be sure that the inactivation process results in a total loss of infectivity.
The use of live, attenuated viral agents as vaccines will often result in improved immunologic reactivity, but increases the risk associated with vaccination in that the vaccine itself is infectious. For example, the virus may revert to an active, virulent form and the organism may be able to propagate and disseminate leading to disease.
Thus, one must generally choose between improved effectiveness and greater degree of safety when selecting between live attenuated and inactivated vaccine preparations. The choice is particularly difficult when the virus is resistant to inactivation processes and requires highly rigorous inactivation conditions which are likely to negatively affect the antigenic characteristics.
Various techniques are known for killing or inactivating viruses for vaccine use. These i

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