New protein engineering approaches to multivalent and bispecific antibody fragments

Plückthun, Andreas; Pack, Peter

doi:10.1016/s1380-2933(97)00067-5

Cited by 296 publications

(158 citation statements)

References 139 publications

Supporting

Mentioning

153

Contrasting

Order By: Relevance

“…In concert with previous studies showing the increased avidity and increased effect of multivalent mAbs, including those based on scFv, Fab, and IgG (13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23), the mAb forms evaluated in this study underscore the potential for enhanced therapeutic utility of multivalent mAbs. Although functionally similar with regard to clustering of target molecules, the multivalent mAb forms vary in size and half-life.…”

Section: Discussionsupporting

confidence: 77%

“…Another tetravalent form was designed by adding Fab at the C terminus of each H chain of a full-length IgG (16). To improve tumor penetrability, smaller constructs using single-chain Fv (scFv) 2 fragments (each Fv consisting of variable light and variable heavy domains connected by peptide linkers) have been joined together to form multivalent complexes (17,18). Such constructs may have relatively short half-lives (compared with those of full-length mAbs), and this has been addressed by joining these scFv multimers to IgG Fc fragments (19,20).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Design, Construction, and In Vitro Analyses of Multivalent Antibodies

Miller¹,

Meng²,

et al. 2003

The Journal of Immunology

View full text Add to dashboard Cite

Some Abs are more efficacious after being cross-linked to form dimers or multimers, presumably as a result of binding to and clustering more surface target to either amplify or diversify cellular signaling. To improve the therapeutic potency of these types of Abs, we designed and generated Abs that express tandem Fab repeats with the aim of mimicking cross-linked Abs. The versatile design of the system enables the creation of a series of multivalent human IgG Ab forms including tetravalent IgG1, tetravalent F(ab′)2, and linear Fab multimers with either three or four consecutively linked Fabs. The multimerized Abs target the cell surface receptors HER2, death receptor 5, and CD20, and are more efficacious than their parent mAbs in triggering antitumor cellular responses, indicating they could be useful both as reagents for study as well as novel therapeutics.

show abstract

Section: Discussionsupporting

confidence: 77%

mentioning

confidence: 99%

Design, Construction, and In Vitro Analyses of Multivalent Antibodies

Miller¹,

Meng²,

et al. 2003

The Journal of Immunology

View full text Add to dashboard Cite

show abstract

“…Because the distance between the carboxyl terminus end of V L and the amino terminus of V H is greater than that for the opposite orientation, it has been discussed that V L -V H orientated scFvs are more constrained than V H -V L oriented fragments when connected by the same linker and therefore tend to exhibit a higher tendency to form higher molecular weight oligomers [30]. Experimental evidence for this assumption has been presented for scFv NC10 by demonstrating that direct linkage of the variable domains in V L -V H orientation resulted in the predominant formation of tetrabodies [9], whereas in reversed orientation formation of trimers was described [10].…”

Section: Discussionmentioning

confidence: 99%

Antigen binding and stability properties of non‐covalently linked anti‐CD22 single‐chain Fv dimers

Arndt¹,

Krauß²,

Rybak³

2004

FEBS Letters

View full text Add to dashboard Cite

By varying linker length and domain orientation three multivalent derivatives of a monovalent anti-CD22 single-chain fragment variable (scFv) antibody were generated. Shortening the linker of the V H -V L oriented scFv to 5 or 0 residues resulted in the formation of diabodies or a mixture of tetramers and trimers, respectively. Unexpectedly, a V L -0-V H scFv assembled to homogenous dimers, remained substantially more stable than the V H -5-V L diabody when incubated in human serum at 37°C, and retained its dimeric state when concentrated up to 4 mg/ml. These properties suggest the V L -0-V H scFv could become an attractive vehicle for the selective delivery of multiple effector molecules to CD22 + tumor cells.

show abstract

“…We expected this molecule to bind with greater avidity to CD123 and CD33 on doublepositive AML cells than the corresponding control sctbs [123 · ds16 · 123] and [33 · ds16 · 33], because it would take advantage of two populations of target antigens rather than only one. Based on theoretical arguments, the avidity of such proteins was expected to be propotional to the combined target antigen density on the tumour cell (Mattes, 1997;Pluckthun & Pack, 1997).…”

mentioning

confidence: 99%

A recombinant trispecific single‐chain Fv derivative directed against CD123 and CD33 mediates effective elimination of acute myeloid leukaemia cells by dual targeting

et al. 2010

View full text Add to dashboard Cite

Two trivalent constructs consisting of single-chain Fv antibody fragments (scFvs) specific for the interleukin-3 receptor a chain (CD123), CD33 and the Fcc-receptor III (CD16) were designed and characterized for the elimination of acute myeloid leukaemia (AML) cells. The dual targeting single-chain Fv triplebody (sctb) [123 · ds16 · 33] and the mono targeting sctb [123 · ds16 · 123] both specifically bound their respective target antigens and were stable in human serum at 37°C for at least 5 d. Both constructs induced potent antibody-dependent cellular cytotoxicity (ADCC) of two different AML-derived CD33-and CD123 double-positive cell lines in the low picomolar range using isolated mononuclear cells (MNCs) as effector cells. In these experiments the dual targeting molecule produced significantly stronger lysis than the mono targeting agent. In addition, the sctbs showed a high potency in mediating ADCC of primary leukaemia cells isolated from peripheral blood or bone marrow of seven AML patients. Hence, these novel molecules displayed potent anti-leukaemic effects against AML cells in vitro and represent attractive candidates for further preclinical development.

show abstract

New protein engineering approaches to multivalent and bispecific antibody fragments

Cited by 296 publications

References 139 publications

Design, Construction, and In Vitro Analyses of Multivalent Antibodies

Design, Construction, and In Vitro Analyses of Multivalent Antibodies

Antigen binding and stability properties of non‐covalently linked anti‐CD22 single‐chain Fv dimers

A recombinant trispecific single‐chain Fv derivative directed against CD123 and CD33 mediates effective elimination of acute myeloid leukaemia cells by dual targeting

Contact Info

Product

Resources

About