Characterization of Noncovalent Complexes of Recombinant Human Monoclonal Antibody and Antigen Using Cation Exchange, Size Exclusion Chromatography, and BIAcore

Santora, Ling; Kaymakcalan, Zehra; Sakorafas, Paul; Krull, Ira S.; Grant, Karen M.

doi:10.1006/abio.2001.5380

Cited by 103 publications

(83 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The dissociation constant (K D ) of TSK114, infliximab and adalimumab for human TNF-α were approximately 5.3 pM, 9.1 nM, and 0.6 nM, respectively (Table 1). The K D values of infliximab and adalimumab determined in this study were higher than the values previously reported, ~0.6 nM of infliximab measured by solid-phase radioimmuno assay (Knight et al, 1993) and ~0.1 nM of adalimumab determined by SPR (Santora et al, 2001). The affinity of TSK114 with human TNF-α was about 1,000-and 100-fold greater than those of infliximab and adalimumab, respectively, mainly as a result of the almost same magnitude reduction in the dissociation constant (k off ) ( Table 1).…”

Section: Kinetic Analysis Of Tsk114 Binding To Human Tnf-αcontrasting

confidence: 88%

Characterization of a novel anti-human TNF-α murine monoclonal antibody with high binding affinity and neutralizing activity

et al. 2008

View full text Add to dashboard Cite

In order to develop an anti-human TNF-α mAb, mice were immunized with recombinant human TNF-α. A murine mAb, TSK114, which showed the highest binding activity for human TNF-α was selected and characterized. TSK114 specifically bound to human TNF-α without cross-reactivity with the homologous murine TNF-α and human TNF-β. TSK114 was found to be of IgG1 isotype with κ light chain. The nucleotide sequences of the variable regions of TSK114 heavy and light chains were determined and analyzed for the usage of gene families for the variable (V), diversity (D), and joining (J) segments. Kinetic analysis of TSK114 binding to human TNF-α by surface plasmon resonance technique revealed a binding affinity (KD) of ~5.3 pM, which is about 1,000-and 100-fold higher than those of clinically relevant infliximab (Remicade) and adalimumab (Humira) mAbs, respectively. TSK114 neutralized human TNF-α-mediated cytotoxicity in proportion to the concentration, exhibiting about 4-fold greater efficiency than those of infliximab and adalimumab in WEHI 164 cells used as an in vitro model system. These results suggest that TSK114 has the potential to be developed into a therapeutic TNF-α-neutralizing antibody with picomolar affinity.

show abstract

Section: Kinetic Analysis Of Tsk114 Binding To Human Tnf-αcontrasting

confidence: 88%

Characterization of a novel anti-human TNF-α murine monoclonal antibody with high binding affinity and neutralizing activity

et al. 2008

View full text Add to dashboard Cite

show abstract

“…Therefore, we decided to study energetics of recognition (stabilization) of native and non-native forms of TNF-␣ by adalimumab, which is the first commercially available therapeutic human antibody. It has been shown that mixing of solutions of adali-mumab and TNF-␣ results in a heterogeneous mixture of TNF-␣-adalimumab complexes (16). Because quantitative thermodynamic analysis and structural modeling of binding events in such a complex interacting system are not possible, we simplified the process by using a fragment of adalimumab (Fab) instead of the full-length antibody.…”

Section: Human Tumor Necrosis Factor ␣ (Tnf-␣) Exists In Its Functionmentioning

confidence: 99%

Recognition of Human Tumor Necrosis Factor α (TNF-α) by Therapeutic Antibody Fragment

Marušič

Podlipnik

Jevševar³

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: Human TNF-␣ is a cytokine involved in many disease-related cellular processes. Results: High affinity binding of therapeutic antibody (inhibitor) to native and molten globule-like TNF-␣ conformation is driven by specific noncovalent interactions. Conclusion: Binding-coupled conformational changes are crucial for antibody-TNF-␣ recognition. Significance: This work helps learn which forces drive unfolding of TNF-␣ and its recognition by monoclonal antibodies and how they affect TNF-␣ activity regulation.

show abstract

“…In addition, monoclonal antibodies are susceptible to chemical or enzymatic modification, particularly at sites that are exposed to the proteinliquid interface. Product heterogeneity can be caused by a number of modifications, such as C-terminal processing of lysine residues (Harris, 1995;Santora et al, 1999;Weitzhandler et al, 1998), deamidation (Di Donato et al, 1993;Hsu et al, 1998), glycation (nonenzymatic glucose addition) (Quan et al, 2008), amino acid sequence variations , and noncovalent complexes (Santora et al, 2001 Schnerman et al, 2004). These orthogonal assays include potency, identity and purity assays, which evaluate "critical quality attributes" such as size and charge heterogeneity.…”

Section: Monoclonal Antibody Characterization and Release Testingmentioning

confidence: 99%

Monoclonal Antibody Development and Physicochemical Characterization by High Performance Ion Exchange Chromatography

Rea¹,

Wang²,

Moreno³

et al. 2012

Innovations in Biotechnology

View full text Add to dashboard Cite

Characterization of Noncovalent Complexes of Recombinant Human Monoclonal Antibody and Antigen Using Cation Exchange, Size Exclusion Chromatography, and BIAcore

Cited by 103 publications

References 15 publications

Characterization of a novel anti-human TNF-α murine monoclonal antibody with high binding affinity and neutralizing activity

Characterization of a novel anti-human TNF-α murine monoclonal antibody with high binding affinity and neutralizing activity

Recognition of Human Tumor Necrosis Factor α (TNF-α) by Therapeutic Antibody Fragment

Monoclonal Antibody Development and Physicochemical Characterization by High Performance Ion Exchange Chromatography

Contact Info

Product

Resources

About