EuroMAbNet: European Monoclonal Antibodies Network

Members

Aurelija Zvirbliene

Name of the laboratory

Members of the laboratory

Unit Leader

Aurelija Zvirbliene azvirb@ibt.lt

Research Fellows

Indre Kucinskaite-Kodze indre.kodze@bti.vu.lt

Indre Dalgediene indre.dalgediene@bti.vu.lt

Dovile Stravinskiene dovile.stravinskiene@gmc.vu.lt

PhD Students

Martynas Simanavicius martynas.simanavicius@bti.vu.lt

Asta Luciunaite asta.luciunaite@gmail.com

Aiste Imbrasaite aiste.imbrasaite@gmc.vu.lt

Laboratory activity

The Dept. of Immunology and Cell Biology is involved in the development and characterization of monoclonal antibodies (MAbs) since 1995. The Department is a part of the newly established Life Sciences Center of Vilnius University with an appropriate infrastructure for cell culture work, immunochemical analysis, molecular biology experiments.

We perform all steps of MAb development - from the generation of recombinant antigens to the detailed characterization of the MAbs including epitope mapping. We also produce recombinant antibodies in different formats.

The network of our international collaborations includes both academic and industrial partners.

Research activities

We have generated hundreds of MAbs against different targets: viral antigens, bacterial virulence factors, cellular proteins, cytokines, hormones, allergens, chemical compounds. The largest antibody collection is generated against viral antigens. These MAbs are valuable tools for investigating antigenic structure of viruses, development of diagnostic assays and prevalence studies of viral infections.

We combine antibody technologies with virus-like particle technologies.

For producing more efficient immunogens, we employ virus-like particles as a carrier for target epitopes to increase their immunogenicity. This approach provides possibilities to generate antibodies against short and non-immunogenic protein sequences.

For construction of recombinant antibodies, we use gene sequences encoding the variable parts of immunoglobulin heavy and light chains from hybridoma cells producing well-characterized MAbs. Recombinant antibodies are generated in different formats - as single-chain antibodies (scFv) and Fc-engineered antibodies where the scFv derived from hybridoma cells is joined to human IgG Fc fragment. Also, we employ recombinant virus-like particles as a carrier for antibody molecules, both scFv and Fc-engineered scFv. This innovative approach allows generation of recombinant multimeric antibodies displayed on virus-like particles.

A significant part of our MAbs are commercialized and/or included into diagnostic kits. We have contracts with industrial partners from Lithuania, UK, Germany, Finland, USA.

Techniques available

• Monoclonal antibody production in mice
• Polyclonal antibody production
• Cloning techniques
• Antibody purification
• Antibody characterization and conjugation
• Epitope mapping
• Mycoplasma testing
• ELISA, Western blot, flow cytometry, immunofluorescence microscopy
• Production of recombinant antigens in bacteria, yeast and mammalian cells
• Construction of chimeric virus-like particles harbouring target epitopes
• Recombinant antibody production
• Construction of recombinant antibodies displayed on virus-like particles

Publications (2019-present)

• Yu JC, Mietzsch M, Singh A, Jimenez Ybargollin A, Kailasan S, Chipman P, Bhattacharya N, Fakhiri J, Grimm D, Kapoor A, Kučinskaitė-Kodzė I, Žvirblienė A, Söderlund-Venermo M, McKenna R, Agbandje-McKenna M. Characterization of the GBoV1 Capsid and Its Antibody Interactions. Viruses. 2021; 13: 330
• Plikusiene I, Maciulis V, Ramanaviciene A, Balevicius Z, Buzavaite-Verteliene E, Ciplys E, Slibinskas R, Simanavicius M, Zvirbliene A, Ramanavicius A. Evaluation of kinetics and thermodynamics of interaction between immobilized SARS-CoV-2 nucleoprotein and specific antibodies by total internal reflection ellipsometry. Journal of Colloid and Interface Science; 2021. 594: 195-203.
• Kučinskaitė-Kodzė I, Simanavičius M, Šimaitis A, Žvirblienė A. Persistence of SARS-CoV-2- Specific Antibodies for 13 Months after Infection. Viruses. 2021.13:2313
• McNabb L, Andiani A, Bulavaite A, Zvirbliene A, Sasnauskas K, Lunt R. Development and validation of an IgM antibody capture ELISA for early detection of Hendra virus. Journal of Virological Methods. 2021; 298:114296
• Rakauskaitė R, Urbanavičiūtė G, Simanavičius M, Žvirblienė A, Klimašauskas S. Selective immunocapture and light-controlled traceless release of transiently caged proteins. STAR Protocols. 2021; 2:100455
• Rakauskaitė Rasa; Urbanavičiūtė Giedrė; Simanavičius Martynas; Lasickienė Rita; Vaitiekaitė Aušra; Petraitytė Gražina; Masevičius Viktoras; Žvirblienė Aurelija; Klimašauskas Saulius. Photocage-Selective Capture and Light-Controlled Release of Target Proteins. iScience. 2020; 23(12): 101833
• Kucinskaite-Kodze I, Simanavicius M, Dapkunas J, Pleckaityte M, Zvirbliene A. Mapping of recognition sites of monoclonal antibodies responsible for the inhibition of pneumolysin functional activity. Biomolecules. 2020; 10, 1009.
• Špakova A, Dalgėdienė I, Insodaitė R, Sasnauskienė A, Žvirblienė A, Petraitytė-Burneikienė R. vB_EcoS_NBD2 bacteriophage-originated polytubes as a carrier for the presentation of foreign sequences. Virus Res. 2020; 290:198194.
• Stravinskiene D, Sliziene A, Baranauskiene L, Petrikaite V, Zvirbliene A. Inhibitory Monoclonal Antibodies and Their Recombinant Derivatives Targeting Surface-Exposed Carbonic Anhydrase XII on Cancer Cells. International Journal of Molecular Sciences. 2020; 21(24): 9411.
• Stravinskiene D, Imbrasaite A, Petrikaite V, Matulis D, Matuliene J, Zvirbliene A. New Monoclonal Antibodies for a Selective Detection of Membrane-Associated and Soluble Forms of Carbonic Anhydrase IX in Human Cell Lines and Biological Samples. Biomolecules. 2019; 9(8): 304

Patents (2010-Present)

• Zvirbliene A., Kucinskaite I., Pleckaityte M., Zvirblis G. Monoclonal antibodies against vaginolysin. EP2398823 (B1), WO2010095917 (A1) (2016).
• Rimseliene, R, Ulcinaite, D, Gagiliene, B, Sukackaite R., Zvirbliene A., Kucinskaite-Kodze I., Dekaminaviciute D. Antibodies that bind thermophillic DNA polymerases. PCT/EP2017/050648 (2016)
• D.Dekaminavičiūtė, M.Plečkaitytė, D.Matulis, A.Žvirblienė. Novel recombinant antibody against human carbonic anhydrase XII. Patent No. LT 6331 B (2016)
• M.Plečkaityte, E.Mištinienė, J.H.Pesliakas, G.Žvirblis, A.Žvirblienė. Recombinant single-chain antibody fragment neutralizing the cytolytic activity of vaginolysin. PCT/LT2011/000004 (2011).
• A.Žvirblienė, A.Gedvilaitė, R.Lasickienė. Monoclonal antibodies against Merkel polyomavirus major capsid protein VP1. Patent LT 5843 B (2011).
• A.Žvirblienė, I.Kučinskaitė-Kodzė. M.Plečkaitytė, G.Žvirblis. Monoclonal antibodies against vaginolysin. Patent LT 5705 B (2010).
• Žvirblienė A., Gedvilaitė A., Ulrich, R., Sasnauskas K. Process for the production of monoclonal antibodies using chimeric VLPs. US 7,919,314 (B2), 2011.