Antitransferrin receptor antibody-RNase fusion protein expressed in the mammary gland of transgenic mice

Newton, Dianne L.; Pollock, David H.; DiTullio, Paul; Echelard, Yann; Harvey, Merri; Wilburn, Brian; Williams, Jennifer L.; Hoogenboom, Hennie R.; Raus, Jef; Meade, Harry; Rybak, Susanna M.

doi:10.1016/s0022-1759(99)00154-4

Cited by 32 publications

(21 citation statements)

References 34 publications

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“…These constructs killed human MDA-MB-231 breast cancer cells, with an IC 50 of 10-40 nmol/l (Newton et al, 1996). A similar cell damaging capability was reported for an anti-TFR mouse-human chimaeric Fab-ANG fusion protein that destroyed different tumour cell lines with IC 50 values ranging from 15 to 70 nmol/l (Newton et al, 1999).…”

Section: Discussionsupporting

confidence: 66%

“…This feature made the soluble bacterial production of a derived humanized scFv-ANG fusion protein impossible. In the past, functional recombinant antibody-ANG fusion proteins were produced from transfected mammalian cells Stocker et al, 2003), recovered from the milk of transgenic animals (Newton et al, 1999), or obtained after production and refolding from insoluble inclusion bodies in bacteria (Newton et al, 1996). As generating transgenic animals or refolding fusion proteins, particularly from insoluble bacterial large scale preparations, is both laborious and expensive, we explored the possibility of producing the humanized fusion protein in mammalian cells.…”

Section: Discussionmentioning

confidence: 99%

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Targeting malignant B‐cell lymphoma with a humanized anti‐CD22 scFv‐angiogenin immunoenzyme‡

Krauss

Arndt

et al. 2005

Br J Haematol

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SummaryWe report on the generation and functional characterization of a humanized immunoenzyme comprising a stable humanized single chain Fv (scFv) with grafted specificity of the anti-CD22 murine monoclonal antibody RFB4 and the human ribonuclease angiogenin (ANG). The fusion protein produced from transiently transfected mammalian Chinese hamster ovary cells could easily be purified to homogeneity, retained full ribonucleolytic activity, and efficiently killed CD22 + tumour cells with an IC 50 of 56 nmol/l. In contrast, incubation of tumour cells with either ANG or scFv alone did not result in any cytotoxicity. Potent receptor-mediated killing of target cells, expected lack of extracellular toxicity, predictable low immunogenic potential, and ease of production, suggest that this novel immunoenzyme has potential for the immunotherapy of CD22 + malignancies.

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Section: Discussionsupporting

confidence: 66%

Section: Discussionmentioning

confidence: 99%

Targeting malignant B‐cell lymphoma with a humanized anti‐CD22 scFv‐angiogenin immunoenzyme‡

Krauss

Arndt

et al. 2005

Br J Haematol

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“…Recombinant human pancreatic ribonuclease A (hPR) was kindly provided by Drs Dianne L. Newton and Susan M. Rybak (SAIC Frederick, NCI at Frederick, National Institutes of Health [NIH]). 24 Human EDN (in this case the Ϫ4 form of EDN that contains 4 additional amino acids on the N-terminus originally considered as part of the signal sequence) 25 was purified from commercial preparations of human urinary gonadotrophin as described. 26 For the production of recombinant mEAR2, the culture supernatants of Spodoptera frugiperda (Sf9) insect cells infected with recombinant baculovirus-encoding mEAR2 were used.…”

Section: Reagentsmentioning

confidence: 99%

Eosinophil-derived neurotoxin (EDN), an antimicrobial protein with chemotactic activities for dendritic cells

Yang

Rosenberg

Chen

et al. 2003

Blood

144

134

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IntroductionAntimicrobial proteins of neutrophil granules have long been considered as important elements of innate host defense. 1 In recent years, evidence has been accumulating indicating that some neutrophil granule-derived antimicrobial proteins also act as chemoattractants for various types of leukocytes. For example, neutrophil-derived ␣-defensins have been shown to be chemotactic for human monocytes, T cells, and immature dendritic cells (iDCs). [2][3][4] Azurocidin and cathelicidins, 2 antimicrobial proteins predominantly stored in neutrophil granules, are chemotactic for neutrophils, monocytes, T cells, and mast cells. 3,[5][6][7][8][9] Cathepsin G, on the other hand, is chemotactic for neutrophils and monocytes. 5 Furthermore, in mouse experimental models, neutrophil granulederived defensins are capable of enhancing antigen-specific immune responses when administered simultaneously with antigens. 10,11 Thus, these neutrophil granule-derived antimicrobial proteins have the capacity to participate in mobilizing and amplifying adaptive immunity by functioning as leukocyte chemoattactants and/or activators. 12 In addition to neutrophil granule-derived antimicrobial proteins, chymase, a serine protease found in the granules of basophils and mast cells, has been shown to induce neutrophil and monocyte chemotaxis. 13 Human eosinophil granules contain several antimicrobial proteins including eosinophil-derived neurotoxin (EDN), 14,15 which together with eosinophil cationic protein belongs to what has come to be recognized as the RNase A superfamily. 16 EDN was purified in 1981 14 and its gene was cloned in 1989. 17 Its 3-dimensional structure has also been solved by X-ray crystallography. 18 Aside from its neurotoxic effect, 14 EDN has been shown to have antiviral activity, in particular against respiratory syncytial virus in vitro, 19,20 suggesting that EDN may contribute to host antiviral defense against single-strand RNA viruses. 21 Very recently, EDN has also been shown to be responsible in part for the HIV-1 inhibitory activities in the supernatant of allogeneic mixed lymphocyte reaction. 22 However, it is not known whether EDN, like several other antimicrobial proteins derived from the granules of neutrophils and basophils/mast cells, may also have chemotactic activity for leukocytes. To test this possibility, we have investigated the capacity of EDN to induce the migration of various types of human leukocytes and found that EDN and its divergent mouse ortholog, mouse eosinophil-associated RNase 2 (mEAR2), 23 can act as selective chemoattractants for DCs. Materials and methods ReagentsRecombinant human (rh) stromal cell-derived factor-1␣ (SDF-1␣), tumor necrosis factor ␣ (TNF␣), granulocyte-macrophage colony-stimulating The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US Government. The publisher or recipient acknowledges right ...

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“…Consequently, several transgenic animals have been developed for the production and expression of human monoclonal and polyclonal antibodies. Mostly, transgenic mice have been used for secretion of the recombinant antibodies 21.3 Outlook 581 into the milk [275][276][277][278]. For the transfer of the expression cassette, chimeric transgenes consisting of the regulatory elements of milk-specific genes and the coding regions of the gene of interest are used.…”

Section: Transgenic Animalsmentioning

confidence: 99%

Emerging Alternative Production Systems

Sommer

Laux

Frenzel

et al. 2014

Handbook of Therapeutic Antibodies

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The market of recombinant protein pharmaceuticals is growing rapidly. Among those, recombinant antibodies are the fastest growing group and a large number of new antibody products are in clinical and preclinical development. Since most conventional cost-effective microbial expression systems are not yet applicable for the expression of fully functional immunoglobulin G (IgG) antibodies, almost all of the currently marketed therapeutic recombinant antibody products are produced from animal cell culture processes that are associated with relative high operating expenses. Thus, to make broad use of recombinant antibody therapeutics more affordable, alternative production systems with improved efficiency and reduced operating expenses are highly desirable.IgG currently is the dominant antibody format for therapeutics. As a heterotetrameric molecule with numerous disulfide bonds and several glycosylation sites IgG makes itself a challenging candidate for heterologous expression, especially in prokaryotes. Glycosylation of the fragment crystallizable (Fc) region has been shown to be essential for mediating effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) [1]. Furthermore, the glycosylation pattern strongly influences the efficiency of the induction of effector functions by antibodies [2, 3]. For applications that depend on the immunological effector functions of the antibody therefore eukaryotic production systems with suitable glycosylation capabilities are necessary.Antibody fragments such as single chain fragment variable (scFv) and fragment antigen binding (Fab) are less complex and the antigen-binding activity is independent of glycosylation, which makes them qualified for prokaryotic expression systems. Certainly, these fragments are incapable of mediating immunological effector functions but for numerous applications this is not necessary or even unwanted.Besides the two traditional expression systems, Escherichia coli for antibody fragments and Chinese hamster ovarian (CHO) or myeloma cells for IgG antibodies, numerous alternative expression systems are currently under development.

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Antitransferrin receptor antibody-RNase fusion protein expressed in the mammary gland of transgenic mice

Cited by 32 publications

References 34 publications

Targeting malignant B‐cell lymphoma with a humanized anti‐CD22 scFv‐angiogenin immunoenzyme‡

Targeting malignant B‐cell lymphoma with a humanized anti‐CD22 scFv‐angiogenin immunoenzyme‡

Eosinophil-derived neurotoxin (EDN), an antimicrobial protein with chemotactic activities for dendritic cells

Emerging Alternative Production Systems

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