Stability and cytotoxicity of Fab-ricin A immunotoxins prepared with water soluble long chain heterobifunctional crosslinking agents

Woo, Byung Ho; Lee, Jung Tae; Park, Myung Ok; Lee, Kang Ro; Han, Jeung Whan; Park, Eun-Seok; Yoo, Sun Dong; Lee, Kang Choon

doi:10.1007/bf02979153

Cited by 8 publications

(6 citation statements)

References 13 publications

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“…The relatively fast cleavage of disulfide linkages in solution was also noted when others prepared disulfide-linked immuno-toxins (i.e., conjugates prepared by conjugating a targeting moiety such as an antibody to a cell-surface molecule and a drug exhibiting cellular toxicity). 26,27 Constrained disulfide linkages were proposed as a means to minimize the linkage cleavage in this case. 26,27 The fast * This analysis relies on an indirect assessment of the extent of thiolBP substitution for HA-bound conjugates.…”

Section: Cleavage Of Hydroxyapatite-bound Conjugatesmentioning

confidence: 99%

“…26,27 Constrained disulfide linkages were proposed as a means to minimize the linkage cleavage in this case. 26,27 The fast * This analysis relies on an indirect assessment of the extent of thiolBP substitution for HA-bound conjugates. The necessary values were obtained from the generated %HA binding data [ Fig.…”

Section: Cleavage Of Hydroxyapatite-bound Conjugatesmentioning

confidence: 99%

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Imparting mineral affinity to proteins with thiol–labile disulfide linkages

Bansal

Wright

Zhang

et al. 2005

J Biomedical Materials Res

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Chemical conjugation of bisphosphonates (BPs) to proteins is an effective means to enhance binding of proteins to mineral-containing biomaterials. BPs linked to proteins with reversible (i.e., cleavable) linkages were considered desirable over the conjugates linked with stable linkages because cleavable linkages allow protein release in free form from the mineral-containing biomaterials. To explore the feasibility of creating cleavable BP-protein conjugates, an amine- and a thiol-containing BP were conjugated to the model protein Bovine Serum Albumin (BSA) with N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), which resulted in disulfide-linked BP-BSA conjugates. Although disulfide-linked conjugates were stable under aqueous conditions, the conjugates in solution were readily cleaved in the presence of physiological concentrations (approximately 0.3 mM) of the thiol compound, cysteine. The imparted mineral affinity as a result of BP conjugation, as assessed by hydroxyapatite (HA) binding in vitro, was lost upon cleavage of the disulfide-linked BP. The conjugates bound to HA were also cleavable with cysteine, but their cleavage rate was significantly reduced as compared to the conjugates in solution. In conclusion, disulfide-linked BP conjugates were shown to be readily cleavable by the amino acid cysteine and this resulted in the loss of imparted mineral affinity of the proteins. The proposed approach will be useful for modulating in vivo delivery of proteins implanted with mineral-containing biomaterials.

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Section: Cleavage Of Hydroxyapatite-bound Conjugatesmentioning

confidence: 99%

Section: Cleavage Of Hydroxyapatite-bound Conjugatesmentioning

confidence: 99%

Imparting mineral affinity to proteins with thiol–labile disulfide linkages

Bansal

Wright

Zhang

et al. 2005

J Biomedical Materials Res

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“…However, as GnRH is a relatively small peptide compared with many large cell recognition molecules such as IgG, GnRH‐cytotoxins may exit the endosome and enter the cytoplasm without being cleaved. Even when large targeting agents are used, evidence is mounting that stable linkers should be considered in conjugate design, including hindered disulfide‐bond linkers, modified short‐chain (Gly‐Phe‐Leu‐Gly) linker, and poly(ethylene glycol)‐dipeptidyl linkers 27–30 . For instance, Cumber et al 31 and Woo et al 29 demonstrated that hindered disulfide‐bonds in immunotoxins had enhanced stability (threefold more stable) compared with non‐hindered disulfide‐bonds.…”

Section: Discussionmentioning

confidence: 99%

A Sandwich Enzyme‐Linked Immunoabsorbent Assay for Measurement of Gonadotropin‐Releasing Hormone‐Toxin Conjugates

Yang

Allen

Wieczorek

et al. 2006

American J Rep Immunol

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“…Clinical trials using immunotoxins have led to studies regarding their optimization focused on both target and toxic domains, although the target domain is the one which has evolved more over the years . Thus, immunotoxins were initially designed containing the entire antibody which soon was replaced by the Fab domain, the single chain variable fragment (scFv) or even solely two antibody complementary determinant regions, aiming to achieve size optimization . Along with these studies, others have focused on the stability of the construct , the use of different coadjuvants or modifications to improve specificity by combining two or more different binding domains to obtain diabodies or triabodies .…”

Section: Introductionmentioning

confidence: 99%

α‐sarcin and RNase T1 based immunoconjugates: the role of intracellular trafficking in cytotoxic efficiency

et al. 2014

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Toxins have been thoroughly studied for their use as therapeutic agents in search of an improvement in toxic efficiency together with a minimization of their undesired side effects. Different studies have shown how toxins can follow different intracellular pathways which are connected with their cytotoxic action inside the cells. The work herein presented describes the different pathways followed by the ribotoxin a-sarcin and the fungal RNase T1, as toxic domains of immunoconjugates with identical binding domain, the single chain variable fragment of a monoclonal antibody raised against the glycoprotein A33. According to the results obtained both immunoconjugates enter the cells via early endosomes and, while a-sarcin can translocate directly into the cytosol to exert its deathly action, RNase T1 follows a pathway that involves lysosomes and the Golgi apparatus. These facts contribute to explaining the different cytotoxicity observed against their targeted cells, and reveal how the innate properties of the toxic domain, apart from its catalytic features, can be a key factor to be considered for immunotoxin optimization.

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Stability and cytotoxicity of Fab-ricin A immunotoxins prepared with water soluble long chain heterobifunctional crosslinking agents

Cited by 8 publications

References 13 publications

Imparting mineral affinity to proteins with thiol–labile disulfide linkages

Imparting mineral affinity to proteins with thiol–labile disulfide linkages

A Sandwich Enzyme‐Linked Immunoabsorbent Assay for Measurement of Gonadotropin‐Releasing Hormone‐Toxin Conjugates

α‐sarcin and RNase T1 based immunoconjugates: the role of intracellular trafficking in cytotoxic efficiency

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