Bispecific antibody generated with sortase and click chemistry has broad antiinfluenza virus activity

Wagner, Kay Uwe; Kwakkenbos, Mark J.; Claassen, Yvonne; Maijoor, Kelly; Böhne, Martino; Sluijs, Koenraad F. van der; Witte, Martin D.; Zoelen, Diana J. van; Cornelissen, Lisette A. H. M.; Beaumont, Tim; Bakker, Arjen Q.; Ploegh, Hidde L.; Spits, Hergen

doi:10.1073/pnas.1408605111

Cited by 70 publications

(75 citation statements)

References 52 publications

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“…Surface plasmon resonance (SPR) analysis was performed on an Ibis Mx96 instrument (Ibis Technologies). Purified antibodies were chemically immobilized on amine-functionalized chips (Ssens, Enschede, The Netherlands) as described before (36).…”

Section: Methodsmentioning

confidence: 99%

Broadly Reactive Anti-Respiratory Syncytial Virus G Antibodies from Exposed Individuals Effectively Inhibit Infection of Primary Airway Epithelial Cells

Cortjens

Yasuda

et al. 2017

J Virol

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Respiratory syncytial virus (RSV) causes severe respiratory disease in young children. Antibodies specific for the RSV prefusion F protein have guided RSV vaccine research, and in human serum, these antibodies contribute to Ͼ90% of the neutralization response; however, detailed insight into the composition of the human B cell repertoire against RSV is still largely unknown. In order to study the B cell repertoire of three healthy donors for specificity against RSV, CD27 ϩ memory B cells were isolated and immortalized using BCL6 and Bcl-xL. Of the circulating memory B cells, 0.35% recognized RSV-A2-infected cells, of which 59% were IgAexpressing cells and 41% were IgG-expressing cells. When we generated monoclonal B cells selected for high binding to RSV-infected cells, 44.5% of IgGexpressing B cells and 56% of IgA-expressing B cells reacted to the F protein, while, unexpectedly, 41.5% of IgG-expressing B cells and 44% of IgA expressing B cells reacted to the G protein. Analysis of the G-specific antibodies revealed that 4 different domains on the G protein were recognized. These epitopes predicted cross-reactivity between RSV strain A (RSV-A) and RSV-B and matched the potency of antibodies to neutralize RSV in HEp-2 cells and in primary epithelial cell cultures. G-specific antibodies were also able to induce antibody-dependent cellular cytotoxicity and antibody-dependent cellular phagocytosis of RSV-A2-infected cells. However, these processes did not seem to depend on a specific epitope. In conclusion, healthy adults harbor a diverse repertoire of RSV glycoprotein-specific antibodies with a broad range of effector functions that likely play an important role in antiviral immunity.IMPORTANCE Human RSV remains the most common cause of severe lower respiratory tract disease in premature babies, young infants, the elderly, and immunocompromised patients and plays an important role in asthma exacerbations. In developing countries, RSV lower respiratory tract disease has a high mortality. Without an effective vaccine, only passive immunization with palivizumab is approved for prophylactic treatment. However, highly potent RSV-specific monoclonal antibodies could potentially serve as a therapeutic treatment and contribute to disease control and mortality reduction. In addition, these antibodies could guide further vaccine development. In this study, we isolated and characterized several novel antibodies directed at the RSV G protein. This information can add to our understanding and treatment of RSV disease.

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Section: Methodsmentioning

confidence: 99%

Broadly Reactive Anti-Respiratory Syncytial Virus G Antibodies from Exposed Individuals Effectively Inhibit Infection of Primary Airway Epithelial Cells

Cortjens

Yasuda

et al. 2017

J Virol

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“…Samples were measured with a FACSCanto or a FACS LSR Fortessa X20 (Becton Dickinson) and analyzed using FlowJo software (Tree Star). The in-house-generated influenza-specific antibody AT1002 (AT10-002) 26 was used as a negative control. To produce recombinant antibody, we isolated total RNA with the RNeasy mini kit (Qiagen), generated cDNA, performed polymerase chain reaction, and cloned the heavy and light chain variable regions into the pCR2.1 TA cloning vector (Invitrogen).…”

Section: Memory Cd27mentioning

confidence: 99%

Patient-derived antibody recognizes a unique CD43 epitope expressed on all AML and has antileukemia activity in mice

et al. 2017

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“…At the same time, it is also possible to tag small molecules with one of the two motifs and use them to produce antibody conjugates. Wagner et al modified LPETG-tagged antibodies with triglycine-functionalized chemical entities that are suitable for copper-free azide-alkyne cycloaddition and subsequently linked two different antibodies to yield bispecific antibody conjugates [129]. Recently, Bellucci et al described a non-canonical function of sortase A where they replaced the polyglycine with a WX3VXVYPKH motif.…”

Section: Sortasementioning

confidence: 99%

Antibody Conjugates: From Heterogeneous Populations to Defined Reagents

2015

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Monoclonal antibodies (mAbs) and their derivatives are currently the fastest growing class of therapeutics. Even if naked antibodies have proven their value as successful biopharmaceuticals, they suffer from some limitations. To overcome suboptimal therapeutic efficacy, immunoglobulins are conjugated with toxic payloads to form antibody drug conjugates (ADCs) and with chelating systems bearing therapeutic radioisotopes to form radioimmunoconjugates (RICs). Besides their therapeutic applications, antibody conjugates are also extensively used for many in vitro assays. A broad variety of methods to functionalize antibodies with various payloads are currently available. The decision as to which conjugation method to use strongly depends on the final purpose of the antibody conjugate. Classical conjugation via amino acid residues is still the most common method to produce antibody conjugates and is suitable for most in vitro applications. In recent years, however, it has become evident that antibody conjugates, which are generated via site-specific conjugation techniques, possess distinct advantages with regard to in vivo properties. Here, we give a comprehensive overview on existing and emerging strategies for the production of covalent and non-covalent antibody conjugates.

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Bispecific antibody generated with sortase and click chemistry has broad antiinfluenza virus activity

Cited by 70 publications

References 52 publications

Broadly Reactive Anti-Respiratory Syncytial Virus G Antibodies from Exposed Individuals Effectively Inhibit Infection of Primary Airway Epithelial Cells

Broadly Reactive Anti-Respiratory Syncytial Virus G Antibodies from Exposed Individuals Effectively Inhibit Infection of Primary Airway Epithelial Cells

Patient-derived antibody recognizes a unique CD43 epitope expressed on all AML and has antileukemia activity in mice

Antibody Conjugates: From Heterogeneous Populations to Defined Reagents

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