Noninvasive brain cancer imaging with a bispecific antibody fragment, generated via click chemistry

Luo, Haiming; Hernandez, Reinier; Hong, Hao; Graves, Stephen A.; Yang, Yunan; England, Christopher G.; Theuer, Charles P.; Nickles, Robert J.; Cai, Weibo

doi:10.1073/pnas.1509667112

Cited by 59 publications

(50 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…165 2 ) by bioorthogonal ''click'' ligation of trans-cyclooctene and tetrazine. 167 Next, the NIR fluorophore ZW800-1 was conjugated to Bs-F(ab) 2 for NIRF imaging of mice bearing U87MG subcutaneous xenografts. ZW800-Bs-F(ab) 2 showed excellent uptake in tumors and low background in non-target tissues.…”

mentioning

confidence: 99%

Fluorescent chemical probes for accurate tumor diagnosis and targeting therapy

et al. 2017

View full text Add to dashboard Cite

Surgical resection of solid tumors is currently the gold standard and preferred therapeutic strategy for cancer. Chemotherapy drugs also make a significant contribution by inhibiting the rapid growth of tumor cells and these two approaches are often combined to enhance treatment efficacy. However, surgery and chemotherapy inevitably lead to severe side effects and high systemic toxicity, which in turn results in poor prognosis. Precision medicine has promoted the development of treatment modalities that are developed to specifically target and kill tumor cells. Advances in in vivo medical imaging for visualizing tumor lesions can aid diagnosis, facilitate surgical resection, investigate therapeutic efficacy, and improve prognosis. In particular, the modality of fluorescence imaging has high specificity and sensitivity and has been utilized for medical imaging. Therefore, there are great opportunities for chemists and physicians to conceive, synthesize, and exploit new chemical probes that can image tumors and release chemotherapy drugs in vivo. This review focuses on small molecular ligand-targeted fluorescent imaging probes and fluorescent theranostics, including their design strategies and applications in clinical tumor treatment. The progress in chemical probes described here suggests that fluorescence imaging is a vital and rapidly developing field for interventional surgical imaging, as well as tumor diagnosis and therapy.

show abstract

mentioning

confidence: 99%

Fluorescent chemical probes for accurate tumor diagnosis and targeting therapy

et al. 2017

View full text Add to dashboard Cite

show abstract

“…Although methods to produce bispecific F(ab') 2 from recombinant Fab' molecules or from human and murine full-length IgG1 have been described previously, 16,8,[17][18][19] no comprehensive method to produce bispecific antibodies directly from all murine IgG isotype antibodies has been published. We described previously a method to produce bispecific F(ab)' 2 from human IgG1 using a bismaleimide crosslinker.…”

Section: Discussionmentioning

confidence: 99%

bisFabs: Tools for rapidly screening hybridoma IgGs for their activities as bispecific antibodies

Patke¹,

Li²,

Wang³

et al. 2017

mAbs

View full text Add to dashboard Cite

Bispecific antibodies are a growing class of therapeutic molecules. Many of the current bispecific formats require DNA engineering to convert the parental monoclonal antibodies into the final bispecific molecules. We describe here a method to generate bispecific molecules from hybridoma IgGs in 3-4 d using chemical conjugation of antigen-binding fragments (Fabs) (bisFabs). Proteolytic digestion conditions for each IgG isotype were analyzed to optimize the yield and quality of the final conjugates. The resulting bisFabs showed no significant amounts of homodimers or aggregates. The predictive value of murine bisFabs was tested by comparing the T-cell redirected cytotoxic activity of a panel of antibodies in either the bisFab or full-length IgG formats. A variety of antigens with different structures and expression levels was used to extend the comparison to a wide range of binding geometries and antigen densities. The activity observed for different murine bisFabs correlated with those observed for the fulllength IgG format across multiple different antigen targets, supporting the use of bisFabs as a screening tool. Our method may also be used for the screening of bispecific antibodies with other mechanisms of action, allowing for a more rapid selection of lead therapeutic candidates.

show abstract

“…The detailed methods for 1,4,7-triazacyclononane-triacetic acid (NOTA) conjugation and 64 Cu labeling with antibodies have been previously reported [14, 15]. Briefly, pertuzumab was mixed with p-SCN-Bn-NOTA in dimethyl sulfoxide (DMSO) and incubated at pH 8.5–9.0 at room temperature for 2 h. NOTA-pertuzumab was purified using PD-10 desalting column with 1 × PBS as the elution buffer.…”

Section: Methodsmentioning

confidence: 99%

Radiolabeled pertuzumab for imaging of human epidermal growth factor receptor 2 expression in ovarian cancer

Jiang

Sun

et al. 2017

Eur J Nucl Med Mol Imaging

Self Cite

View full text Add to dashboard Cite

Purpose Human epidermal growth factor receptor 2 (HER2) is overexpressed in over 30% of ovarian cancer cases, playing essential roles in tumorigenesis and metastasis. Non-invasive imaging of HER2 is of great interest for physicians to better detect and monitor the progression of ovarian cancer. In this study, HER2 was assessed as a biomarker for ovarian cancer imaging using 64Cu-labeled pertuzumab for immunoPET imaging. Methods HER2 expression and binding were examined in three ovarian cancer cell lines (SKOV3, OVCAR3, Caov3) using in vitro techniques, including Western blot and saturation binding assays. PET imaging and biodistribution studies in subcutaneous models of ovarian cancer were performed to non-invasively evaluate HER2 expression in vivo. Additionally, orthotopic models were employed to further validate the imaging capability of 64Cu-NOTA-pertuzumab. Results HER2 expression was highest in SKOV3 cells, while OVCAR3 and Caov3 displayed lower HER2 expression. 64Cu-NOTA-pertuzumab showed high specificity to HER2 (Ka = 3.1 ± 0.6 nM) in SKOV3. In subcutaneous tumors, PET imaging revealed tumor uptakes of 41.8 ± 3.8, 10.5 ± 3.9, and 12.1 ± 2.3 %ID/g at 48 h post-injection for SKOV3, OVCAR3, and Caov3, respectively (n=3). In orthotopic models, PET imaging with 64Cu-NOTA-pertuzumab allowed for rapid and clear delineation of both primary and small peritoneal metastases in HER2-overexpressing ovarian cancer. Conclusion 64Cu-NOTA-pertuzumab is an effective PET tracer for the non-invasive imaging of HER2-expression in vivo, making it a potential tracer for treatment monitoring and improved patient stratification in the future.

show abstract

Noninvasive brain cancer imaging with a bispecific antibody fragment, generated via click chemistry

Cited by 59 publications

References 43 publications

Fluorescent chemical probes for accurate tumor diagnosis and targeting therapy

Fluorescent chemical probes for accurate tumor diagnosis and targeting therapy

bisFabs: Tools for rapidly screening hybridoma IgGs for their activities as bispecific antibodies

Radiolabeled pertuzumab for imaging of human epidermal growth factor receptor 2 expression in ovarian cancer

Contact Info

Product

Resources

About