US 9,394,339 B2  20  19  protect against enzymatic deamidation during production  and/or storage ofa vaccine formulation. If the interpretation  above is reasonably correct, then a complex or intermittent  covalent bond between stabilizer and labile site would also be  expected to limit enzymatic deamidation, by a sort of com- petitive inhibition at the labile site. Enzymatic deamidation is  believed to occur in certain hepatitis vaccines, for example.  Hemoglobin is known to undergo nonenzymatic deamida› tion, and while it is not a traditional active pharmaceutical  ingredient, it is being investigated as a key ingredient in  artificial blood (blood substitute), and thus formulations con› taining hemoglobin or related heme protein for blood substi› tutes are within the scope ofthis invention.  Embodiments of the current invention can be used for  many routes of administration compatible with peptide or  protein delivery, including but not limited to parenteral, intra› venous, intraperitoneal, intrathecal, intramuscular, subcuta› neous,  intra-arterial,  rectal,  intravaginal,  sublingual,  intraocular, transdermal, intranasal, via inhalation, in a sup› pository, ophthalmic, and others. Oral delivery of peptides  and proteins is currently a largely unsolved problem,  although several promising approaches have made progress,  and the invention may be compatible with some such formu› lations.  It should be noted that while the primary function of the  current invention is to minimize or prevent deamidation,  other degradation reactions occurring in the formulations of  the invention might also be concomitantly reduced by the  stabilizing anion(s). This is particularly true for other base› catalyzed reactions involving carbonyl groups.  A deamidation-prone decapeptide with two labile aspar› aginyl residues was shown in this Example to be stabilized  against deamidation by the current invention; the Example  also shows the destabilizing effect of anions such as acetate  and phosphate. To begin with, a decapeptide was synthesized  by request at the Protein Facility of the Iowa State University  Office of Biotechnology, using standard peptide synthesis  methodology. The peptide had the sequence RCNSGR› FNGG-NH2 (Arg-Cys-Asn-Ser-Gly-Arg-Phe-Asn-Gly-Gly,  terminal amide). Literature on peptide degradation by deami› dation at asparagine sites shows by overwhelming consensus  that the FNG and CNS subsequences are prone to degrada› tion.  Samples containing this decapeptide were prepared under  eight different conditions at two different pH values, pH 7 and  4. The 200 mM arginine/I 00 mM phosphate buffer was only  prepared at pH 7. The preparation of these samples is now  described. The ratios indicated are molar ratios of organic  anion to decapeptide.  HCI-Control.  Into a sterile 15 mL plastic centrifuge tube (Coming Incor› porated, Coming, N.Y.) was placed 0.0021 g decapeptide  (Iowa State University, Ames, Iowa) and 7.0 g sterile water  (Spectrum Chemicals Mfg. Corp., Gardena, Calif.). The pH  was adjusted using I .ON HCl (Spectrum Chemicals Mfg.  Corp., Gardena, Calif.)to a pH of 4. Into anew 15 mL plastic  centrifuge tube was placed 3 .5 mL of a pH 4 solution. The pH  was adjusted using only the necessary amount of I .ON NaOH  (Spectrum Chemicals Mfg. Corp., Gardena, Calif.) to a final  pHof7.  Saccharin:Decapeptide, 1: 1  The potency (mg/mL of intact peptide) was determined by  integrating under this peak (performed by the instrument),  and normalizing according to a standard run of the decapep› tide. The relative potency was then calculated by dividing the  Chromatographic analysis.  The samples at the various timepoints were then analyzed  with HPLC using the following conditions.  System: Schimadzu SCL-l0A VP with diode array detector  Colunm: Phenomenex C18 Luna 5 um, 250x4.6 mm  Mo bile Phase: Isocratic 90% ( 0 .1 % ) TF A in water, 10%  (0.08%) TFA in acetonitrile  Flow Rate: 1.0 mL/min  Wavelength: 214 nm  Into a sterile 1.5 mL microcentrifuge tube was placed a 0.5  g portion of each sample condition and were placed at -18°  C.  This was the initial sample. The remaining pH 4 samples were  placed at 65°  C. until completion of the experiment. The  remaining pH 7 samples were placed at 40°  C. until comple› tion of the experiment. Into a sterile 1.5 mL microcentrifuge  tube was placed a 0.5 g portion of each sample condition on  "day 1", 24 hours after preparation, and were placed at -18°  C. This was the day 1 sample. The frozen initial and day 1  samples were thawed to room temperature, and assayed using  HPLC.  Into a sterile 15 mL plastic centrifuge tube was placed  0.0021 g decapeptide, 0.0016 g sodium acetate, 0.0094 g  sodium saccharin and 7 .0 g sterile water. The pH was adjusted  using only the necessary amount of acid saccharin to a pH of  4. Into a new 15 mL plastic centrifuge tube was placed 3 .5 mL  of a pH 4 solution. The pH was adjusted using only the  necessary amount of I .ON Na OH to a final pH of 7.  Acetate/Saccharin:Decapeptide, 3/10: 1.  Into a sterile 15 mL plastic centrifuge tube was placed  0.0021 g decapeptide, 0.0016 g sodium acetate (Spectrum  Chemicals Mfg. Corp., Gardena, Calif.) and 7.0 g sterile  water. The pH was adjusted using only the necessary amount  of acetic acid (Spectrum Chemicals Mfg. Corp., Gardena,  Calif.) to a pH of 4. Into a new 15 mL plastic centrifuge tube  was placed 3.5 mL of a pH 4 solution. The pH was adjusted  using only the necessary amount of I .ON NaOH to a final pH  of7.  Acetate:Decapeptide, 3: 1.  Into a 15 mL plastic centrifuge tube was placed 0.0021 g  decapeptide, 0.0095 g sodium saccharin and 7.0 g sterile  water. The pH was adjusted using only the necessary amount  of acid saccharin to a pH of 4. Into a new 15 mL plastic  centrifuge tube was placed 3 .5 mL of a pH 4 solution. The pH  was adjusted using only the necessary amount of 1.0NNaOH  toafinalpHof7.  Saccharin:Decapeptide, 10: 1.  Into a sterile 15 mL plastic centrifuge tube was placed  0.0021 g decapeptide, 0.0029 g sodium saccharin and 7.0 g  sterile water. The pH was adjusted using only the necessary  amount of saccharin to a pH of 4. Into a new 15 mL plastic  centrifuge tube was placed 3 .5 mL of a pH 4 solution. The pH  was adjusted using only the necessary amount of 1.0NNaOH  to a final pH of 7.  Saccharin:Decapeptide, 3: 1  ( anion:asparaginyl molar ratio). Into a sterile 15 mL plastic  centrifuge tube was placed 0.0021 g decapeptide, 0.0011 g  sodium saccharin (Spectrum Chemicals Mfg. Corp., Gar› dena, Calif.) and 7.0 g sterile water. The pH was adjusted  using only the necessary amount of saccharin (Spectrum  Chemicals Mfg. Corp., Gardena, Calif.) to a pH of 4. Into a  new 15 mL plastic centrifuge tube was placed 3 .5 mL of a pH  4 solution. The pH was adjusted using only the necessary  amount of I .ON Na OH to a final pH of 7.  EXAMPLES  Example 1 