Insulin, separation

Insulin glulisine Insulin Separate CV and respiratory No, but the 12-month Standard Recombinant FDA-2004... 

Insulin glargine Insulin Separate safety pharmacolgy. 2 year (mouse) Standard Recombinant FDA-2000... 

Design, Treatment and Sample Size This was a single-dose, randomized, open-label, two-way, crossover study. 0.1 lU/kg insulin glulisine and 0.2 lU/kg NPH insulin separate and simultaneous s.c. injections in the abdominal area or 0.1 lU/kg insulin glulisine and 0.2 lU/kg NPH insulin by s.c. injection in the abdominal area, immediately after being premixed in the syringe. 

Dimov, N. Simeonov, S. Experimental models for optimization of insulin separation on reversed phase columns. Biomed.Chromatogr, 1994, 8, 32—36 [gradient cow pig]... 

A search was made for optimal conditions of insulin separation by comparing three different silica-based chromatographic supports in GPC under denaturing conditions [112]. The separation patterns were identical with those obtained with soft gels. 

The concentration of insulin in a production vat is determined by the method of isotope dilution. A 1.00-mg sample of insulin labeled with with an activity of 549 cpm, is added to a 10.0-mL sample taken from the production vat. After homogenizing the sample, a portion of the insulin is separated and purified, yielding 18.3 mg of pure insulin. The activity for the isolated insulin is measured at 148 cpm. How many milligrams of insulin are in the original sample ... 

Human insulin was the first animal protein to be made in bacteria in a sequence identical to the human pancreatic peptide. Expression of separate insulin A and B chains were achieved in Echerichia coli K-12 using genes for the insulin A and B chains synthesized and cloned in frame with the... 

Fig. 5. Anion-exchange separation of insulin and insulin A- and B-chains, over diethylaminoethyl (DEAF) in a 10.9 x 200 mm column having a volume of 18.7 mL. Sample volume is 0.5 mL and protein concentration ia 16.7 mAf Tris buffer at pH 7.3 is 1 mg/mL for each component ia the presence of EDTA. Eluent (also 16.7 mAf Tris buffer, pH 7.3) flow rate is 1.27 ml,/min, and protein detection is by uv absorbance at 280 nm. The straight line depicts the salt...

This reversed-phase chromatography method was successfully used in a production-scale system to purify recombinant insulin. The insulin purified by reversed-phase chromatography has a biological potency equal to that obtained from a conventional system employing ion-exchange and size-exclusion chromatographies (14). The reversed-phase separation was, however, followed by a size-exclusion step to remove the acetonitrile eluent from the final product (12,14). 

It is often important to control the CSD of pharmaceutical compounds, eg, in the synthesis of human insulin, which is made by recombinant DNA techniques (1). The most favored size distribution is one that is monodisperse, ie, all crystals are of the same size, so that the rate at which the crystals dissolve and are taken up by the body is known and reproducible. Such uniformity can be achieved by screening or otherwise separating the desired size from a broader distribution or by devising a crystallization process that will produce insulin in the desired form. The latter of these options is preferable, and considerable effort has been expended in that regard. 

Analogous to two-dimensional LC, the on-line coupling of LC and CE has been carried out both in the heart-cut and the comprehensive mode. In a heart-cut LC-CE study, a protein G immunoaffinity LC column was used to selectively preconcentrate insulin from serum (171). A 1 p.1 elution plug comprising the insulin was switched on-line to the CE system where a part was injected into the capillary for final separation. With CE, efficient separations can be obtained in a... 

An unexpected response may be obtained when changing from mixed injections to separate injections or vice versa If the patient had been using insulin mixtures before admission, the nurse asks whether the insulins were given separately or together. 

Human insulin Two peptide chains A, 21 amino acids long, and B, 30 amino acids long . coli Juvenile onset diabetes Approved for sale A and B chains made separately as fusion proteins and joined in vitro Compared with animal Insulins some undesirable side-effects have been noted... 

Kasicka, V., Pruslk, Z., Sazelova, P., Jiracek, J. and Barth, T., Theory of the correlation between capillary and free-flow zone electrophoresis and its use for the conversion of analytical capillary separations to continuous free-flow preparative processes. Application to analysis and preparation of fragments of insulin, ]. Chromatogr. A, 796, 211, 1998. 

CF-related diabetes shares characteristics of both type 1 and type 2 diabetes mellitus but is categorized separately. The primary cause of CFRD is insulin deficiency resulting from both reduced functional pancreatic islet cells and increased islet amyloid deposition. Insulin secretion is delayed in response to glucose challenge, and absolute insulin secretion over time is reduced. Some insulin resistance may also be present in CFRD however, sensitivity may be increased in CF patients without diabetes.8... 

Insulin is the one agent that can be used in all forms of DM for blood sugar control. Insulin is the essential treatment for patients with type 1 DM and can overcome insulin resistance in patients with type 2 DM. Insulin is available commercially in various formulations that vary markedly in terms of onset and duration of action and the source from which a product is obtained. Insulins can be divided into four separate classes based on their length of action. Most formulations are available as U-100, indicating a concentration of 100 units/mL. Insulin is typically refrigerated, and most vials are good for 28 days at room temperature. Specific details of insulin products are listed in Table 40-9. 

A number of combination insulin products are available commercially. NPH is available in combinations of 70/30 and 50/50 with regular insulin. Two short-acting insulin analog mixtures are also available. Humalog Mix 75/25 contains 75% insulin lispro protamine suspension and 25% insulin lispro. Novolog Mix 70/30 contains 70% insulin aspart protamine suspension and 30% insulin aspart. The lispro and aspart insulin protamine suspensions were developed specifically for these mixture products and will not be commercially available separately. 

Online LC-ESI-TOF-MS experiments are carried out in a very similar fashion to the off-line NPS-HPLC separations described above, with a few notable exceptions. Firstly, 0.3% (v/v) formic acid is added to each mobile phase to counteract the ionization suppression induced by TFA. Because of the formic acid UV detection must be carried out at 280 nm (as opposed to 214 nm). To aid in normalization between runs 1 jag of Bovine insulin (MW = 5734 Da) is added to each chromatofocusing fraction prior to injection onto the column. Finally, the flow is split postcolumn directing 200 JlL/min into the ion source and the remaining 300 JlL/min through the UV detector and fraction collection. 

As described above all samples were separated online using LCT ESI-TOF-MS then normalized for relative quantitation using a bovine insulin internal standard. Fractions were then collected for MAFDI-TOF-MS PMF, digested with modified porcine trypsin, and analyzed using the TofSpec2E. Following this analysis, three major classes of differentially expressed including proteins were revealed in these... 

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