Stabilization and formulation of a recombinant Human Cytomegalovirus vector for use as a candidate HIV-1 vaccine

Ozan S. Kumru, Soraia Saleh-Birdjandi, Lorena R. Antunez, Eddy Sayeed, David Robinson, Sjoerd van den Worm, Geoffrey S. Diemer, Wilma Perez, Patrizia Caposio, Klaus Früh, Sangeeta B. Joshi, David B. Volkin

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Live attenuated viral vaccine/vector candidates are inherently unstable and infectivity titer losses can readily occur without defining appropriate formulations, storage conditions and clinical handling practices. During initial process development of a candidate vaccine against HIV-1 using a recombinant Human Cytomegalovirus vector (rHCMV-1), large vector titer losses were observed after storage at 4 °C and after undergoing freeze-thaw. Thus, the goal of this work was to develop candidate frozen liquid formulations of rHCMV-1 with improved freeze-thaw and short-term liquid stability for potential use in early clinical trials. To this end, a virus stability screening protocol was developed including use of a rapid, in vitro cell-based immunofluorescence focus assay to quantitate viral titers. A library of ∼50 pharmaceutical excipients (from various known classes of additives) were evaluated for their effect on vector stability after freeze-thaw cycling or incubation at 4 °C for several days. Certain additives including sugars and polymers (e.g., trehalose, sucrose, sorbitol, hydrolyzed gelatin, dextran 40) as well as removal of NaCl (lower ionic strength) protected rHCMV-1 against freeze-thaw mediated losses in viral titers. Optimized solution conditions (e.g., solution pH, buffers and sugar type) slowed the rate of rHCMV-1 titer losses in the liquid state at 4 °C. After evaluating various excipient combinations, three new candidate formulations were designed and rHCMV-1 stability was benchmarked against both the currently-used and a previously reported formulation. The new candidate formulations were significantly more stable in terms of reducing rHCMV-1 titer losses after 5 freeze-thaw cycles or incubation at 4 °C for 30 days. This case study highlights the utility of semi-empirical design of frozen liquid formulations of a live viral vaccine candidate, where protection against infectivity titer losses due to freeze-thaw and short-term liquid storage are sufficient to enable more rapid initiation of early clinical trials.

Original languageEnglish (US)
Pages (from-to)6696-6706
Number of pages11
JournalVaccine
Volume37
Issue number44
DOIs
StatePublished - Oct 16 2019

Fingerprint

Viral Vaccines
Human herpesvirus 5
AIDS Vaccines
Excipients
Human immunodeficiency virus 1
Cytomegalovirus
HIV-1
Clinical Trials
vaccines
Attenuated Vaccines
Trehalose
liquids
Sorbitol
Gelatin
Dextrans
Osmolar Concentration
Fluorescent Antibody Technique
Sucrose
live vaccines
Buffers

Keywords

  • Cytomegalovirus
  • Excipient
  • Formulation
  • Freeze-thaw
  • HIV vaccine
  • Stability

ASJC Scopus subject areas

  • Molecular Medicine
  • Immunology and Microbiology(all)
  • veterinary(all)
  • Public Health, Environmental and Occupational Health
  • Infectious Diseases

Cite this

Kumru, O. S., Saleh-Birdjandi, S., Antunez, L. R., Sayeed, E., Robinson, D., van den Worm, S., ... Volkin, D. B. (2019). Stabilization and formulation of a recombinant Human Cytomegalovirus vector for use as a candidate HIV-1 vaccine. Vaccine, 37(44), 6696-6706. https://doi.org/10.1016/j.vaccine.2019.09.027

Stabilization and formulation of a recombinant Human Cytomegalovirus vector for use as a candidate HIV-1 vaccine. / Kumru, Ozan S.; Saleh-Birdjandi, Soraia; Antunez, Lorena R.; Sayeed, Eddy; Robinson, David; van den Worm, Sjoerd; Diemer, Geoffrey S.; Perez, Wilma; Caposio, Patrizia; Früh, Klaus; Joshi, Sangeeta B.; Volkin, David B.

In: Vaccine, Vol. 37, No. 44, 16.10.2019, p. 6696-6706.

Research output: Contribution to journalArticle

Kumru, OS, Saleh-Birdjandi, S, Antunez, LR, Sayeed, E, Robinson, D, van den Worm, S, Diemer, GS, Perez, W, Caposio, P, Früh, K, Joshi, SB & Volkin, DB 2019, 'Stabilization and formulation of a recombinant Human Cytomegalovirus vector for use as a candidate HIV-1 vaccine', Vaccine, vol. 37, no. 44, pp. 6696-6706. https://doi.org/10.1016/j.vaccine.2019.09.027
Kumru OS, Saleh-Birdjandi S, Antunez LR, Sayeed E, Robinson D, van den Worm S et al. Stabilization and formulation of a recombinant Human Cytomegalovirus vector for use as a candidate HIV-1 vaccine. Vaccine. 2019 Oct 16;37(44):6696-6706. https://doi.org/10.1016/j.vaccine.2019.09.027
Kumru, Ozan S. ; Saleh-Birdjandi, Soraia ; Antunez, Lorena R. ; Sayeed, Eddy ; Robinson, David ; van den Worm, Sjoerd ; Diemer, Geoffrey S. ; Perez, Wilma ; Caposio, Patrizia ; Früh, Klaus ; Joshi, Sangeeta B. ; Volkin, David B. / Stabilization and formulation of a recombinant Human Cytomegalovirus vector for use as a candidate HIV-1 vaccine. In: Vaccine. 2019 ; Vol. 37, No. 44. pp. 6696-6706.
@article{3be401428986440393dc3a66ab065dfc,
title = "Stabilization and formulation of a recombinant Human Cytomegalovirus vector for use as a candidate HIV-1 vaccine",
abstract = "Live attenuated viral vaccine/vector candidates are inherently unstable and infectivity titer losses can readily occur without defining appropriate formulations, storage conditions and clinical handling practices. During initial process development of a candidate vaccine against HIV-1 using a recombinant Human Cytomegalovirus vector (rHCMV-1), large vector titer losses were observed after storage at 4 °C and after undergoing freeze-thaw. Thus, the goal of this work was to develop candidate frozen liquid formulations of rHCMV-1 with improved freeze-thaw and short-term liquid stability for potential use in early clinical trials. To this end, a virus stability screening protocol was developed including use of a rapid, in vitro cell-based immunofluorescence focus assay to quantitate viral titers. A library of ∼50 pharmaceutical excipients (from various known classes of additives) were evaluated for their effect on vector stability after freeze-thaw cycling or incubation at 4 °C for several days. Certain additives including sugars and polymers (e.g., trehalose, sucrose, sorbitol, hydrolyzed gelatin, dextran 40) as well as removal of NaCl (lower ionic strength) protected rHCMV-1 against freeze-thaw mediated losses in viral titers. Optimized solution conditions (e.g., solution pH, buffers and sugar type) slowed the rate of rHCMV-1 titer losses in the liquid state at 4 °C. After evaluating various excipient combinations, three new candidate formulations were designed and rHCMV-1 stability was benchmarked against both the currently-used and a previously reported formulation. The new candidate formulations were significantly more stable in terms of reducing rHCMV-1 titer losses after 5 freeze-thaw cycles or incubation at 4 °C for 30 days. This case study highlights the utility of semi-empirical design of frozen liquid formulations of a live viral vaccine candidate, where protection against infectivity titer losses due to freeze-thaw and short-term liquid storage are sufficient to enable more rapid initiation of early clinical trials.",
keywords = "Cytomegalovirus, Excipient, Formulation, Freeze-thaw, HIV vaccine, Stability",
author = "Kumru, {Ozan S.} and Soraia Saleh-Birdjandi and Antunez, {Lorena R.} and Eddy Sayeed and David Robinson and {van den Worm}, Sjoerd and Diemer, {Geoffrey S.} and Wilma Perez and Patrizia Caposio and Klaus Fr{\"u}h and Joshi, {Sangeeta B.} and Volkin, {David B.}",
year = "2019",
month = "10",
day = "16",
doi = "10.1016/j.vaccine.2019.09.027",
language = "English (US)",
volume = "37",
pages = "6696--6706",
journal = "Vaccine",
issn = "0264-410X",
publisher = "Elsevier BV",
number = "44",

}

TY - JOUR

T1 - Stabilization and formulation of a recombinant Human Cytomegalovirus vector for use as a candidate HIV-1 vaccine

AU - Kumru, Ozan S.

AU - Saleh-Birdjandi, Soraia

AU - Antunez, Lorena R.

AU - Sayeed, Eddy

AU - Robinson, David

AU - van den Worm, Sjoerd

AU - Diemer, Geoffrey S.

AU - Perez, Wilma

AU - Caposio, Patrizia

AU - Früh, Klaus

AU - Joshi, Sangeeta B.

AU - Volkin, David B.

PY - 2019/10/16

Y1 - 2019/10/16

N2 - Live attenuated viral vaccine/vector candidates are inherently unstable and infectivity titer losses can readily occur without defining appropriate formulations, storage conditions and clinical handling practices. During initial process development of a candidate vaccine against HIV-1 using a recombinant Human Cytomegalovirus vector (rHCMV-1), large vector titer losses were observed after storage at 4 °C and after undergoing freeze-thaw. Thus, the goal of this work was to develop candidate frozen liquid formulations of rHCMV-1 with improved freeze-thaw and short-term liquid stability for potential use in early clinical trials. To this end, a virus stability screening protocol was developed including use of a rapid, in vitro cell-based immunofluorescence focus assay to quantitate viral titers. A library of ∼50 pharmaceutical excipients (from various known classes of additives) were evaluated for their effect on vector stability after freeze-thaw cycling or incubation at 4 °C for several days. Certain additives including sugars and polymers (e.g., trehalose, sucrose, sorbitol, hydrolyzed gelatin, dextran 40) as well as removal of NaCl (lower ionic strength) protected rHCMV-1 against freeze-thaw mediated losses in viral titers. Optimized solution conditions (e.g., solution pH, buffers and sugar type) slowed the rate of rHCMV-1 titer losses in the liquid state at 4 °C. After evaluating various excipient combinations, three new candidate formulations were designed and rHCMV-1 stability was benchmarked against both the currently-used and a previously reported formulation. The new candidate formulations were significantly more stable in terms of reducing rHCMV-1 titer losses after 5 freeze-thaw cycles or incubation at 4 °C for 30 days. This case study highlights the utility of semi-empirical design of frozen liquid formulations of a live viral vaccine candidate, where protection against infectivity titer losses due to freeze-thaw and short-term liquid storage are sufficient to enable more rapid initiation of early clinical trials.

AB - Live attenuated viral vaccine/vector candidates are inherently unstable and infectivity titer losses can readily occur without defining appropriate formulations, storage conditions and clinical handling practices. During initial process development of a candidate vaccine against HIV-1 using a recombinant Human Cytomegalovirus vector (rHCMV-1), large vector titer losses were observed after storage at 4 °C and after undergoing freeze-thaw. Thus, the goal of this work was to develop candidate frozen liquid formulations of rHCMV-1 with improved freeze-thaw and short-term liquid stability for potential use in early clinical trials. To this end, a virus stability screening protocol was developed including use of a rapid, in vitro cell-based immunofluorescence focus assay to quantitate viral titers. A library of ∼50 pharmaceutical excipients (from various known classes of additives) were evaluated for their effect on vector stability after freeze-thaw cycling or incubation at 4 °C for several days. Certain additives including sugars and polymers (e.g., trehalose, sucrose, sorbitol, hydrolyzed gelatin, dextran 40) as well as removal of NaCl (lower ionic strength) protected rHCMV-1 against freeze-thaw mediated losses in viral titers. Optimized solution conditions (e.g., solution pH, buffers and sugar type) slowed the rate of rHCMV-1 titer losses in the liquid state at 4 °C. After evaluating various excipient combinations, three new candidate formulations were designed and rHCMV-1 stability was benchmarked against both the currently-used and a previously reported formulation. The new candidate formulations were significantly more stable in terms of reducing rHCMV-1 titer losses after 5 freeze-thaw cycles or incubation at 4 °C for 30 days. This case study highlights the utility of semi-empirical design of frozen liquid formulations of a live viral vaccine candidate, where protection against infectivity titer losses due to freeze-thaw and short-term liquid storage are sufficient to enable more rapid initiation of early clinical trials.

KW - Cytomegalovirus

KW - Excipient

KW - Formulation

KW - Freeze-thaw

KW - HIV vaccine

KW - Stability

UR - http://www.scopus.com/inward/record.url?scp=85072535951&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85072535951&partnerID=8YFLogxK

U2 - 10.1016/j.vaccine.2019.09.027

DO - 10.1016/j.vaccine.2019.09.027

M3 - Article

C2 - 31548012

AN - SCOPUS:85072535951

VL - 37

SP - 6696

EP - 6706

JO - Vaccine

JF - Vaccine

SN - 0264-410X

IS - 44

ER -