US 2020/0246423 Al  73  Aug. 6, 2020  TABLE 57  TABLE 59  TABLE 60  TABLE 58  Concentration, pH, and Osmolality  [0418]  The aflibercept concentration (mg/mL), pH, and  osmolality (mOSm) of the Block E formulations were  evaluated at tO. Results are reported in Table 59 below.  [0419]  The stability of the Block E formulations was  evaluated by measuring the amount of non-degraded afliber› cept present in the formulation at time  O  (tO) and after being  subjected to storage conditions. Size Exclusion Chromatog› raphy was conducted on the formulations and the percentage  of protein present in the main peak (non-degraded afliber› cept) in each formulation is reported in Table 60-63 below.  Size Exclusion Chromatography  [0417]  This example describes experiments to evaluate  buffer-free aflibercept formulations that have a stabilizer  other than sucrose (Block E). The design mirrored Block D,  except all of these compositions include 0.03% PS 20. This  will allow for a direct and detailed examination of the  importance PS 20 (and co-solvents in general) for their  impact on storage stability. Table 58, below, shows an  exemplary set of such formulations.  [0416]  It  is further contemplated that in the formulations  of Block D, phosphate could be used instead of histidine  buffer.  It  is also contemplated that CaC1$_{2 }$ could be used  instead of, or in addition to, MgC1$_{2}$ .  [0415]  The aflibercept formulations of Block Dare physi› cally and chemically stable.  Discussion  Example 6-Buffer-Free Aflibercept Formulations  with Alternative Stabilizers  ic!EF results for Block D formulations after storage for four weeks at 5°  C.  Rel. Area (%) Smoothing (Grou12)  Formulation Block  2  3  4  5  6  7  8  9  D-1  4.6 6.8 7.4  11.4  13.9  15.2  14.5  11.6  8.5  D-2  4.5 6.2 7.8  12.1  14.6  14.9  14.2  11.6  8.0  D-3  4.5 6.1  7.7  11.9  14.2  14.8  14.5  11.6  8.5  D-4  4.3 6.6 7.3  12.4  14.0  15.5  14.1  11.5  8.5  D-5  4.8 6.3 7.3  12.5  13.9  14.7  14.9  11.6  8.2  D-6  4.2 7.1  6.9  11.6  14.8  15.0  14.2  11.8  8.1  D-7  4.4 6.3  8.0  12.0  14.3  14.8  14.7  11.6  8.0  D-8  3.9 6.5 7.2  12.1  15.1  15.0  14.1  11.2  8.3  D-9  4.2 6.7 7.3  12.1  14.3  15.7  14.1  11.8  8.0  D-10  4.4 6.1  8.0  11.9  14.4  15.6  14.0  11.9  7.8  D-11  4.8 6.8 7.4  12.3  13.8  15.7  14.1  11.6  7.9  D-12  5.0 6.5  6.7  12.4  14.7  15.4  14.1  11.9  7.8  Eylea  4.5 6.2 7.8  12.1  14.6  14.9  14.2  11.6  8.0  10  11  12  3.9 1.8 0.4  3.8 1.8 0.5  3.6 1.9 0.7  3.6 1.8 0.5  3.5 1.9 0.5  3.5 1.9 0.9  3.6  1.7  0.5  3.7 2.1  0.8  3.7 1.6 0.4  3.8 1.6 0.4  3.6 1.6 0.3  3.5 1.6 0.4  3.8 1.6 0.6  Form  Final Concentration  Final Osmolality  No.  (mg/mL)  Final pH  (mOsm/Kg)  E-1  38.33  6.23  274  E-2  37.88  6.21  303  E-3  39.55  6.25  284  E-4  40.06  6.27  242  E-5  37.91  6.14  295  E-6  39.63  6.11  271  E-7  39.31  6.22  287  E-8  39.85  6.12  290  E-9  39.57  6.44  276  E-10  42.04  6.57  297  E-11  39.71  6.95  276  E-12  40.42  6.94  302  All formulations include 0.03% PS 20.  Form  Gly  Arg  Pro  NaCl  MgC1$_{2 }$ Dextran  No.  aflibercept pH (mM) (mM) (mM) (mM)  (mM)  (%)  E-1  40 mg/ml  6.2  270  0  0  0  0  0  E-2  40 mg/ml  6.2  0  0  270  0  0  0  E-3  40 mg/ml  6.2  0  25  0  125  0  0  E-4  40 mg/ml  6.2  0  50  0  75  0  0  E-5  40 mg/ml  6.2  0  0  0  150  0  E-6  40 mg/ml  6.2  0  0  0  0  100  0  E-7  40 mg/ml  6.2  270  0  0  0  0  E-8  40 mg/ml  6.2  0  0  0  40  75  0  E-9  40 mg/ml  6.5  270  0  0  0  0  0  E-10  40 mg/ml  6.5  0  0  270  0  0  0  E-11  40 mg/ml 7.0  270  0  0  0  0  0  E-12  40 mg/ml 7.0  0  0  270  0  0  0  SEC results for Block E formulations at T - 0  Form  Rel. Area(%),  Rel. Area (%),  Rel. Area (% ),  No.  Before Main Peak  Main Peak  After Main Peak  E-1  1.11  98.89  0  E-2  1.14  98.86  0  E-3  1.14  98.86  0  E-4  1.14  98.86  0  E-5  1.18  98.82  0  E-6  1.43  98.57  0  E-7  1.15  98.85  0  E-8  1.09  98.91  0  E-9  1.13  98.87  0  E-10  1.19  98.81  0  E-11  1.29  98.71  0  E-12  1.36  98.64  0 