Optimization on Critical Parameters for Lyophilization of Reference Standard Viral Vaccines by Using Freeze-Drying Microscopy

Main Article Content

Assajun Amen
Pattama Bunnag
Kornnika Kullabutr

Abstract

Background: Freeze-drying or lyophilization is a process that preserves a suitable qualified for standard reference viral vaccine to control a viral vaccine quality. The exploration on real lyophilization state need the use of lots of vaccine samples and spent approximately 1-5 days a time, also it tested several times to get a suitable state.


Objective: The aim of this research was to develop a providing standard reference viral vaccine by using a freeze-drying microscopy that observed a changing of the samples while simulation, temperature, and pressure of lyophilization to verify a collapse temperature.


Methods: The study set four formulas to test the collapse temperature that under freeze drying microscopy, and took the temperature average that subsidence to calculate a primary freeze-drying temperature to set the state of lyophilization machine. Then, it trailed the real lyophilization and took these samples to test an appearance, potency, and stability.


Results: The primary drying temperatures of four formulas, that calculated, were -27.8, -28.8, -29.0, and -28.8 OC, respectively. The difference of all four formulas was not statistically significant. After taking the standards reference viral vaccines from the lyophilization to test an appearance; moisture content; reconstitution time; and percentage of potency lost (before and after lyophilization), it is found that four formulas had good characteristic and were on criteria of each testing. It excepted the test of stability at 37 OC in seven days that discovered the highest level of virus potency lost by 1.54 log PFU/0.5 ml.


Conclusion: The lyophilization method of standard reference viral vaccine by using freeze-drying microscopy was suitable for setting primary drying temperature. It was able to control temperature that was not too high until the samples collapsed and was not too low that caused using more time for lyophilization. It used the test time only 0.5-2 hours and used less samples at microliter level. Consequently, this method is suitable and able to use the country standard of freeze-drying parameter finding.

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Section
Research Article
Author Biographies

Assajun Amen, Institute of Biological Products, Department of Medical Sciences

Background: Freeze-drying or lyophilization is a process that preserves a suitable qualified for standard reference viral vaccine to control a viral vaccine quality. The exploration on real lyophilization state need the use of lots of vaccine samples and spent approximately 1-5 days a time, also it tested several times to get a suitable state.

Objective: The aim of this research was to develop a providing standard reference viral vaccine by using a freeze-drying microscopy that observed a changing of the samples while simulation, temperature, and pressure of lyophilization to verify a collapse temperature.

Methods: The study set four formulas to test the collapse temperature that under freeze drying microscopy, and took the temperature average that subsidence to calculate a primary freeze-drying temperature to set the state of lyophilization machine. Then, it trailed the real lyophilization and took these samples to test an appearance, potency, and stability.

Results: The primary drying temperatures of four formulas, that calculated, were -27.8, -28.8, -29.0, and -28.8 OC, respectively. The difference of all four formulas was not statistically significant. After taking the standards reference viral vaccines from the lyophilization to test an appearance; moisture content; reconstitution time; and percentage of potency lost (before and after lyophilization), it is found that four formulas had good characteristic and were on criteria of each testing. It excepted the test of stability at 37 OC in seven days that discovered the highest level of virus potency lost by 1.54 log PFU/0.5 ml.

Conclusion: The lyophilization method of standard reference viral vaccine by using freeze-drying microscopy was suitable for setting primary drying temperature. It was able to control temperature that was not too high until the samples collapsed and was not too low that caused using more time for lyophilization. It used the test time only 0.5-2 hours and used less samples at microliter level. Consequently, this method is suitable and able to use the country standard of freeze-drying parameter finding.

Pattama Bunnag, Institute of Biological Products, Department of Medical Sciences

Background: Freeze-drying or lyophilization is a process that preserves a suitable qualified for standard reference viral vaccine to control a viral vaccine quality. The exploration on real lyophilization state need the use of lots of vaccine samples and spent approximately 1-5 days a time, also it tested several times to get a suitable state.

Objective: The aim of this research was to develop a providing standard reference viral vaccine by using a freeze-drying microscopy that observed a changing of the samples while simulation, temperature, and pressure of lyophilization to verify a collapse temperature.

Methods: The study set four formulas to test the collapse temperature that under freeze drying microscopy, and took the temperature average that subsidence to calculate a primary freeze-drying temperature to set the state of lyophilization machine. Then, it trailed the real lyophilization and took these samples to test an appearance, potency, and stability.

Results: The primary drying temperatures of four formulas, that calculated, were -27.8, -28.8, -29.0, and -28.8 OC, respectively. The difference of all four formulas was not statistically significant. After taking the standards reference viral vaccines from the lyophilization to test an appearance; moisture content; reconstitution time; and percentage of potency lost (before and after lyophilization), it is found that four formulas had good characteristic and were on criteria of each testing. It excepted the test of stability at 37 OC in seven days that discovered the highest level of virus potency lost by 1.54 log PFU/0.5 ml.

Conclusion: The lyophilization method of standard reference viral vaccine by using freeze-drying microscopy was suitable for setting primary drying temperature. It was able to control temperature that was not too high until the samples collapsed and was not too low that caused using more time for lyophilization. It used the test time only 0.5-2 hours and used less samples at microliter level. Consequently, this method is suitable and able to use the country standard of freeze-drying parameter finding.

References

Rey L, May JC. Freeze-drying/ lyophilization of pharmaceutical and biological products, 2nd ed. NewYork: Marcel Dekker; 2004.

Singh SK, Upadhyay RC, Yadav MC, Tiwari M. Development of a novel lyophilization protocol for preservation of mushroom mycelial cultures, Curr Sci [Internet]. 2004 [cited 2021 Apr 23]; 87(5):568-70. Available from: https://www.researchgate.net/profile/Dr-Mahesh-Yadav/publication/282819934_Development_of_a_novel_lyophilization_protocol_for_preservation_of_mushroom_mycelial_cultures/links/5697300308ae34f3cf1e1022/Development-of-a-novel-lyophilization-protocol-for-preservation-of-mushroom-mycelial-cultures.pdf

ปัณณธร ภัทรสถาพรกุล. เทคโนโลยีการทำแห้งแบบเยือกแข็ง (ตอนที่ 3). วารสารสมาคมเครื่องทำความเย็นไทย [อินเทอร์เน็ต]. 2548 [เข้าถึงเมื่อ 23 เม.ย. 2564];15:7-9. เข้าถึงได้จาก: https://tra.or.th/wp-content/uploads/2017/05/keep-kool-15.pdf

Ohtomo T, Yamada T, Yoshida K. Outermost-cell-surface changes in an encapsulated strain of Staphylococcus aureus after preservation by freeze-drying. Appl Environ Microbiol [Internet]. 1988 [cited 2021 Apr 23]; 54(10):2486-91. Available from: https://aem.asm.org/content/aem/54/10/2486.full.pdf

Naddafi K, Moosavi GH, Mesdaghinia AR. Evaluation of lyophilization effects on operational parameters and characteristics of activated sludge. Iranian J Env Health Sci Eng [Internet]. 2004 [cited 2021 Apr 23]; 1(1):20-5. Available from:https://www.sid.ir/FileServer/JE/102620040109.pdf

Jennings TA. Lyophilization: introduction and basic principles. NewYork: Informa Healthcare USA; 2008.

อัศจรรย์ อาเมน, จิรเดช ปัจฉิม, พรหมฉัตร เจริญพัฒน์, ณัฐกานต์ มิ่งงามทรัพย์, สุภาพร ภูมิอมร. การศึกษาหาสูตรตำรับที่เหมาะสมในการทำแห้งวัคซีนคางทูมอ้างอิงมาตรฐาน. วารสารกรมวิทยาศาสตร์การแพทย์. 2554;53(2):97-112.

Zhai S, Taylor R, Sanches R. Slater NKH. Measurement of lyophilisation primary drying rates by freeze-drying microscopy. Chem Eng Sci 2003;58(11):2313–23.

Kevin R. Advanced uses of freeze drying microscopy (fdm) for product formulation and lyo-cycle development [Internet]. 2007 [cited 2019 Sep 1]. Available from: http://docplayer.net/39803625-Advanced-uses-of-freeze-drying-microscopy-fdm-for-product-formulation-and-lyo-cycle-development.html

Barresi AA, Ghio S, Fissore D, Pisano R. Freeze‐drying of pharmaceutical excipients close to collapse temperature: influence of the process conditions on process time and product quality. Drying Technology 2009;27(6):805-16.

Ross C, Gaster T, Ward K. The Importance of Critical Temperatures in the Freeze Drying of Pharmaceutical Products [Internet]. 2011 [cited 2019 Sep 1]. Available from: http://biopharma.co.uk/intelligent-freeze-drying/wp-content/uploads/sites/2/2011/07/Importance_critical_temps_Resized-v1.pdf

Kasraian K, Spitznagelb TM, Juneau J, Yim K. Characterization of the Sucrose/Glycine/ Water System by Differential Scanning Calorimetry and Freeze-Drying Microscopy, Pharmaceut Dev Tech 1998;3(2):233-9.

Tang X, Pikal MJ. Design of freeze-drying processes for pharmaceuticals: practical advice. Pharmaceut Res 2004;21:191–200.

Lu, X, Pikal M. Freeze-drying of mannitol-trehalose-sodium chloride-based formulations: The impact of annealing on dry layer resistance to mass transfer and cake structure. Pharmaceut Dev Tech 2004;9:85–95.

Haeusera C, Goldbach P, Huwyler J, Friess W, Allmendinger A. Impact of dextran on thermal properties, product quality attributes, and monoclonal antibody stability in freeze-dried formulations. Eur. J. Pharm. Biopharm 2020;24(147):45-56.

Meister E, Gieseler H. Freeze-Dry Microscopy of Protein/Sugar Mixtures: Drying Behavior, Interpretation of Collapse Temperatures and a Comparison to Corresponding Glass Transition Data. J Pharmaceut Sci 2009;98(9):3072-87.

Pikal MJ, Shah S. The collapse temperature in freeze drying: Dependence on measurement methodology and rate of water removal from the glassy phase. Int J Pharm 1990;62:165-86.

Rambhatla S, Obert JP, Luthra S, Bhugra C, Pikal MJ. Cake shrinkage during freeze drying: A combined experimental and theoretical study. Pharmaceut Dev Tech 2005;10(1):33-40.

WHO Expert Committee on Biological Standardization. Annex 7: Recommendations to assure the quality, safety and efficacy of Japanese encephalitis vaccines (live, attenuated) for human use [Internet]. 2014 [cited 2019 Sep 1]. Available from: https://www.who.int/biologicals/vaccines/JE-Recommendations_TRS_980_

Annex_7.pdf?ua=1

ณัฐกานต์ มิ่งงามทรัพย์, อัศจรรย์ อาเมน, สุภาพร ภูมิอมร. การทำแห้งไวรัสโรตาซีโรทัยป์เดี่ยวเชื้อเป็น เพื่อใช้เป็นวัคซีนอ้างอิงมาตรฐานของประเทศ. วารสารวิชาการสาธารณสุข 2559;25(5):897-907.