DNA production

BollonA.P. (l9S4)Recombinant DNA Products Insulin, Interferon andGrowth Hormone. Boca Raton, Florida CRC Press. 

Biotechnology era beginning First recombinant DNA products Human insulin Human growth hormone Interferons, etc. Monoclonal antibodies Nucleotide blockage Growth in use of natural products and neutraceuticals... 

Biomolecular MS and in particular MALDI-TOF-MS (see Sections 2.1.22 and 2.2.1) permit the routine analysis of oligonucleotides up to 70-mers, intact nucleic acids, and the direct detection of DNA products with no primer labels with an increase in analysis speed and mass accuracy especially in contrast to traditional DNA separation techniques such as slab gels or capillary electrophoresis. Applications focus on the characterization of single nucleotide polymorphisms (SNPs) and short tandem repeats (STRs). Precise and accurate gene expression measurements show relative and absolute numbers of target molecules determined independently of the number of PCR cycles. DNA methylation can be studied quantitatively. 

Stebbing, N. and Week, P.K. (1984). Preclinical assessment of biological properties of recombinant DNA-derived human interferons. In Recombinant DNA Products Insulin, Interferon and Growth Hormone, (Bollon, A.P., Ed.). CRC Press, Boca Raton, FL. 

In the late 1970s, development of recombinant DNA products utilizing knowledge of cellular and molecular biology commenced. The biotechnology industry became a reality. 

In some ways, the use of animals (almost always mammals) as substitutes for bacteria in the recombinant DNA production of drugs is a natural and obvious extension of the techniques originally developed for the manufacture of insulin, human growth hormone, and other pharmaceuticals. Live animals have a built-in production... 

Nitrous oxide exerts a variety of its adverse effects by oxidizing vitamin Bn and rendering it inactive as a coenzyme in many essential metabolic processes. One vitamin dependent enzyme in particular, methionine synthetase, is involved in cell division and is necessary for DNA production. Adverse reproductive and hematologic effects caused by nitrous oxide are thought to be due to inactivation or dysfunction of methionine synthetase resulting in impairment of cell division. 

Antibodies to hydroxymethyl uracil, an oxdized DNA base, were determined in workers exposed to nickel and cadmium, and in welders (Frenkel et al. 1994). Compared to controls, a significant increase in these antibodies was noted in the most highly exposed workers. Personal monitoring of 12 workers exposed to nickel and cadmium showed correlation coefficients between exposure concentrations and the antibodies of 0.4699 for cadmium and 0.7225 for nickel. Antibodies to hydroxymethyl uracil were not increased among welders. The levels of antibodies in the control populations for the nickel cadmium workers and for the welders were different indicating the importance of determining the distribution of a new biomarker in controls for each population that is studied. This preliminary study suggests that antibodies to oxidized DNA products may be useful biomarkers for nickel and other metals that induce oxidative stress. 

Although increases in oxidized DNA products and precancerous and cancerous lesions in the nose have been associated with niekel exposure, these effects are not speeilie to niekel. There are no specific biomarkers for niekel health efifeets. 

Interferon alfa-2b (Intron A) is a recombinant DNA product derived from the interferon alfa-2b gene of human white blood cells. Its mechanism of antitumor action involves binding to a plasma membrane receptor but is otherwise poorly understood. Its serum half-life is 2 to 3 hours after parenteral administration. 

RNA polymerase, and the triphosphates of the purine ribonucleosides, uridine, and cytidine, and otherwise the same conditions, will prime the synthesis of RNA. The amounts of synthetic DNA or RNA are many fold greater than the amount of primer DNA the DNA product is nearly the same in most measurable ways as the primer DNA. The efficiency of the DNA in initiating these syntheses is known as the primer activity of the DNA, and can be affected by alterations of the bases which compose the nucleic acid, and by other factors. 

In 1996, about 10 years after the introduction of the first recombinant DNA product for human use, the FDA modified and streamlined the approval process for biotechnology products considered to be well characterized. These modifications, in essence, established the direction of how biologic macromolecules are researched and developed today in biotechnology-based and traditional pharmaceutical companies [2]. Well-characterized biotechnology products include (1) synthetic peptides consisting of fewer than 20 amino acids, (2) monoclonal antibodies and derivatives, and (3) recombinant DNA-derived products. Anticipating future developments, the FDA is also prepared to consider DNA plasmid products as well-characterized when the first medicinal in this class is submitted for approval. CBER now approves well-characterized biopharmaceuticals under the BLA process [3]. 

Roeder, T., Simple and efficient cloning of small polymerase chain reaction-generated DNA products. Anal Biochem, 2000.285(2) 278-80. 

PaUvizumab, humanized monoclonal IgGi against F protein of respiratory syncytial virus (RSV), recombinant DNA product... 

Commercial insulin preparations differ in a number of ways, such as differences in the recombinant DNA production techniques, amino acid sequence, concentration, solubility, and the time of onset and duration of their biologic action. 

With chromatin suspensions, DNA product yields are an order of magnitude lower as compared to DNA solutions. This is partially due to OH-scavenging by the nucleoproteins, but since one deals here with suspensions, OH-scavenging by dissolved impurities has an even more dramatic effect than in DNA solutions. Moreover, some of the DNA damage maybe repaired by the surrounding protein, but the existing data do not provide a firm conclusion concerning this point. 

N(6)-furfuryladenine (kinetin) is formed in vivo, excreted in the urine, and has been assumed to be a secondary, free-radical-induced DNA product (Bar-ciszewski et al. 1996,1997,1999,2000 Wyszko et al. 2003). 

Table 12.9. Kinetic isotope effects for the formation of some DNA products connected with DNA strand breakage by OH. (Balasubramanian et al. 1998)...

An important contributor to DNA-protein cross-links maybe the reaction of DNA products with proteins. A case in point is the cross-linking of the relatively abundant DNA damage 2-dRL with a lysine moiety [reaction (39) Hashimoto et al., 2001]. 

Delahoussaye YM, Hay MP, Pruijn FB, Denny WA, Brown JM (2003) Improved potency of the hypoxic cytotoxin tirapazamine by DNA targeting. Biochem Pharmacol 65 1807-1815 Demple B, Linn S (1982) 5,6-Saturated thymine lesions in DNA production by UV light or hydrogen peroxide. Nucleic Acids Res 10 3781-3789... 

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