Investigation into alternative testing methodologies for characterization of influenza virus vaccine

Figures & data

Table 1. Details of examined lots of bioCSL's IVV for the 2013 SH season

Lot no.	No. of syringes
090634903	18
090635001	18
090636401	18
090637301	18
090638202	18

Table 2. Results of description test and additional comments for 5 lots of bioCSL's 2013 SH IVV (18 syringes per lot, 90 syringes in total)

IVV lot no.	Description test	Comments test
090634903	18 out of 18 PASS	All syringes were identified to contain “small amounts of fine sediment observed in vaccine, all sediment resuspended upon shaking.”
090636401	18 out of 18 PASS
090638202	18 out of 18 PASS
090637301	18 out of 18 PASS	From each lot, 15 syringes were identified to contain “small amounts of fine sediment” and the remaining 3 syringes comprised “large amounts and/or large sized particles of sediment,” all of which “resuspended upon shaking.”
090635001	18 out of 18 PASS

Figure 1. Representative ILM images for 5 lots of bioCSL's 2013 SH IVV, including lots 090634903, 090635001, 090636401, 090637301 and 090638202, when syringes were left unshaken (left column) and following vigorous shaking (right column).

Table 3. Summary of ILM results for the appearance of bioCSL's 2013 SH IVV

Lot no.	No. of syringes	Unshaken (US) / Shaken (S)	Observations: morphology of sediment, if present, is consistent with that of influenza vaccine?
090634903	15	US	YES
		S	YES
090635001	15	US	YES
		S	YES
090636401	15	US	YES
		S	YES
090637301	15	US	YES
		S	YES
090638202	15	US	YES
		S	YES

Figure 2. Representative EM images for pooled material corresponding to each of the 5 lots of bioCSL's 2013 SH IVV at a magnification of 108,500x for lots (A) 090634903, (B) 090635001, (C) 090636401, (D) 090637301 and (E) 090638202; and at a magnification of 32,200x for lot (F) 090638202.

Figure 3. Average protein recovery in supernatant (expressed as %ODT) for 5 pooled groups within each of 5 lots of bioCSL's 2013 SH IVV, including lots (A) 090634903, (B) 090635001, (C) 090636401, (D) 090637301 and (E) 090638202, by ODT analysis post-centrifugation at an RCF of 13,684 g for 1 min.

Table 4. Density, viscosity and refractive index (RI) properties of pooled 2013 SH IVV for DLS analysis at 25°C

	Pooled 2013 SH IVV
Density (g/cm^3)	1.003
Viscosity (cP)	0.923
RI	1.3342

Figure 4. Average intensity PSDs (n = 5) of the supernatant for 5 pooled groups within each of 5 lots of bioCSL's 2013 SH IVV, including lots (A) 090634903, (B) 090635001, (C) 090636401, (D) 090637301 and (E) 090638202, by DLS analysis at 25°C post-centrifugation at an RCF of 6,082 g for 1 min.

Figure 5. Average volume PSDs (n = 5) of the supernatant for 5 pooled groups within each of 5 lots of bioCSL's 2013 SH IVV, including lots (A) 090634903, (B) 090635001, (C) 090636401, (D) 090637301 and (E) 090638202, by DLS analysis at 25°C post-centrifugation at an RCF of 6,082 g for 1 min.

Figure 6. Average UV-visible absorbance (AU) vs. wavelength (nm) profiles for 5 lots of bioCSL's 2013 SH IVV, including lots (A) 090634903, (B) 090635001, (C) 090636401, (D) 090637301 and (E) 090638202, performed neat for each lot (n = 3).

Table 5. Agglomeration index (AI;%) for 5 lots of 2013 SH IVV, determined from UV-visible absorbance spectroscopy analysis

2013 SH IVV Lot no.	Average A280	Average A350	AI (%)
090634903	0.430	0.136	46.2
090635001	0.430	0.135	45.6
090636401	0.479	0.155	47.6
090637301	0.444	0.142	46.9
090638202	0.415	0.138	49.6

Table 6. Example ODT analysis for IVV

	Initial A280 (mg/ml) [A]	Supernatant post-centrifugation A280 (Sample 1) (mg/ml) [B]	Supernatant post-centrifugation A280 (Sample 2) (mg/ml) [C]	% Recovery (Sample 1) [D]	% Recovery (Sample 2) [E]	Average % Recovery [F]	Standard Deviation (stdev) of % Recovery (±)
Description of analysis	A280 of neat sample (2 replicates)	A280 of supernatant after centrifugation (2 replicates)	A280 of supernatant after centrifugation (2 replicates)	= B / A * 100	= C / A * 100	= (D + E) / 2 * 100	= stdev (D and E)
Example IVV	Avg A280 (0.61, 0.59) = 0.60	Avg A280 (0.36, 0.35) = 0.36	Avg A280 (0.36, 0.33) = 0.35	59.2	57.5	58.3	1.18