Typing reagents for hybrid glycophorin blood groups that are rare in Caucasians, but with prevalence of up to 8% in East Asian ethnic groups, are unavailable despite repeated attempts by various groups to use conventional approaches to produce useful mAbs. We will utilise naive human phage libraries to develop reagents that react with RBC with a phenotype defined by the most common hybrid glycophorin but not with pooled human RBC that do not display these antigens. The usefulness of the mAb produced as a typing reagent will be assessed by binding to the target hybrid glycophorin detected by flow cytometry. Subsequently reactivity using the standard immunohaematological agglutination based techniques will be assessed as well as characterising other features required for typing reagents, such as long term stability at room temperature.  

Lead investigator	Dr Martina Jones
Postdoc	Dr Xuan Bui
Industry Partner	ARCBS; PIs, Flower and Irving (ARCBS)

Integral membrane proteins are attractive targets for research, diagnostic and therapeutic applications, as they act as biomarkers to define a particular cell type, developmental stage or disease type. As such, mAbs against cell-surface biomarkers are highly sought after as biological therapeutics, laboratory reagents or diagnostic reagents. Based on antibody phage display methodologies.6 AIBN has developed novel whole-cell biopanning techniques to improve the efficiency of screening antibody libraries on whole cells displaying biomarkers. The proposed collaborative project aims to pool the skills of AIBN and CSL researchers to further optimise the whole cell biopanning technique to isolate new mAbs against specific cell surface biomarkers that are of interest to CSL. The outcomes of this project would be further innovations in whole cell panning methodologies, as well as isolation of new antibodies of therapeutic significance.

Lead investigator	Dr Martina Jones
Postdoc	Dr Sumukh Kumble
Industry Partner	CSL; PI, Panousis (CSL)

Using N-ethyl-N-nitrosourea (ENU)-induced mutagenesis, ARCBS has identified a unique murine pedigree with a splice-site mutation in a functionally important domain of a gene encoding an acetyltransferase. Red Blood Cells (RBC) from homozygous mutants from this pedigree demonstrate significant modifications of carbohydrates and lack the erythroid lineage marker TER-119. We plan to investigate the extent to which these changes modify the susceptibility to infection for pathogens that target developing and mature cells of the erythroid lineage. We will compare RBC from homozygous and wild type (WT) mice to characterise the RBC surface and identify novel RBC targets to be used as a basis for the development of biopharmaceuticals for treatment of diseases where infectivity is mediated through RBC receptors (e.g. Malaria, Parvovirus (B19)).

Lead investigator	Prof Stephen Mahler
Postdoc	Dr Xuan Bui
Industry Partner	ARCBS; PIs, Dean, Flower and Irving (ARCBS)

Human neutrophil antigens (HNAs) are a group of 5 glycoproteins that are expressed on human neutrophils, and in some cases, also on other cells and tissues. Endogenous antibodies against HNAs have been implicated in cases of alloimmune neonatal neutropenias, autoimmune neutropenias, febrile transfusion reactions and transfusion-related acute lung injury (TRALI). The availability of mAbs to HNAs permits the use of solid phase assays to detect endogenous antibodies specific to these antigens. While mAb reagents against HNA-1, HNA-2, HNA-4 and HNA-5 antigens are available, there are not yet any mAbs available for HNA-3. This project aims to develop mAbs against the two alleles of HNA-3 that can then be used to develop cell-based assays to detect anti-HNA-3a and anti-HNA-3b antibodies in blood donors. Furthermore, this project aims to use the anti-HNA-3 mAbs in existing in vitro transfusion models to help understand the mechanisms by which TRALI develops.

Lead investigator	Dr Martina Jones
Postdoc	Dr Xuan Bui
Industry Partner	ARCBS; PIs, Tung, Flower and Irving (ARCBS)

Intracellular processes including protein transport, transcription and signalling present a range of new potential targets that could have application in the treatment of a variety of disease indications. The screening of chemical libraries for binders to intracellular targets has had limited success due to (i) the relative lack of binding clefts and hydrophobic pockets compared to those located on cell surface proteins and (ii) the small “footprint” to which a small molecule can bind on an extensive protein surface interface. As an alternative, antibody fragments are being explored as molecular entities that are capable of disrupting protein-protein interactions within the cell, traditionally thought to be undruggable.5 This project will investigate strategies for intracellular delivery of antibodies and antibody fragments, along with assessing the resulting impact on cellular processes. An antibody against the transcription factor SOX18 (created at AIBN and IMB) will be used as the initial model system for intracellular antibody delivery. A successful outcome of this project will be the establishment of platform technology for the delivery of antibody fragments to intracellular targets.  

Project contacts

Project Leader	Dr Mathias Francois
Postdoc	Dr Sumukh Kumble
Industry Partner	CSL