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Factors influencing the quality of Myrmecia pilosula (Jack Jumper) ant venom for use in in vitro and in vivo diagnosis of allergen sensitization and in allergen immunotherapy

Citation

Wanandy, T and Dwyer, HE and McLean, L and Davies, NW and Nichols, D and Gueven, N and Brown, SGA and Wiese, MD, Factors influencing the quality of Myrmecia pilosula (Jack Jumper) ant venom for use in in vitro and in vivo diagnosis of allergen sensitization and in allergen immunotherapy, Clinical and Experimental Allergy, 47, (11) pp. 1478-1490. ISSN 1365-2222 (2017) [Refereed Article]

Copyright Statement

ę 2017 John Wiley & Sons Ltd

DOI: doi:10.1111/cea.12987

Abstract

Background

Allergen immunotherapy uses pharmaceutical preparations derived from naturally occurring source materials, which contain water-soluble allergenic components responsible for allergic reactions. The success of inávivo and inávitro diagnoses in allergen sensitization and allergen immunotherapy largely depends on the quality, composition and uniformity of allergenic materials used to produce the active ingredients, and the formulation employed to prepare finished products.

Objectives

We aimed to examine the factors influencing batch-to-batch consistency of Jack Jumper (Myrmecia pilosula) ant venom (JJAV) in the form of active pharmaceutical ingredient (AI) and informed whether factors such as temperature, artificial light and container materials influence the quality of JJAV AIs. We also aimed to establish handling and storage requirements of JJAV AIs to ensure preservation of allergenic activities during usage in the diagnosis of allergen sensitization and in allergen immunotherapy.

Methods

The quality and consistency of JJAV AIs were analysed using a combination of bicinchoninic acid assay for total protein quantification, HPLC-UV for JJAV allergen peptides quantification, ELISA inhibition for total allergenic potency, SDS-PAGE, AU-PAGE and immunoblot for qualitative assessment of JJAV components, and Limulus Amebocyte Lysate assay for the quantification of endotoxin concentration. API-ZYM and Zymogram assays were used to probe the presence of enzymatic activities in JJAV.

Results

Pharmaceutical-grade JJAV for allergen immunotherapy has good batch-to-batch consistency. Temporary storage at 4░C and light exposure do not affect the quality of JJAV. Exposure to temperature above 40░C degrades high MW allergens in JJAV. Vials containing JJAV must be stored frozen and in upright position during long-term storage.

Conclusions and Clinical Relevance

We have identified factors, which can influence the quality and consistency of JJAV AIs, and provided a framework for appropriate handling, transporting and storage of JJAV to be used for the diagnosis of allergen sensitization and in AIT.

Item Details

Item Type:Refereed Article
Keywords:jack jumper, venom, allergy, immunotherapry
Research Division:Medical and Health Sciences
Research Group:Immunology
Research Field:Allergy
Objective Division:Health
Objective Group:Clinical Health (Organs, Diseases and Abnormal Conditions)
Objective Field:Immune System and Allergy
Author:Wanandy, T (Mr Troy Wanandy)
Author:Dwyer, HE (Ms Hayley Dwyer)
Author:McLean, L (Mrs Lee McLean)
Author:Davies, NW (Associate Professor Noel Davies)
Author:Nichols, D (Dr David Nichols)
Author:Gueven, N (Associate Professor Nuri Gueven)
Author:Brown, SGA (Professor Simon Brown)
Author:Wiese, MD (Associate Professor Michael Wiese)
ID Code:120179
Year Published:2017
Deposited By:Central Science Laboratory
Deposited On:2017-08-14
Last Modified:2017-11-20
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