Engineering of Affibody Molecules for Therapy and Diagnostics

2012 ◽  
pp. 103-126 ◽  
Author(s):  
Joachim Feldwisch ◽  
Vladimir Tolmachev
Keyword(s):  
Affibody Molecules

Specific Targeting of HER2-Positive Head and Neck Squamous Cell Carcinoma Line HN5 by Idarubicin-ZHER2 Affibody Conjugate

2018 ◽  
Vol 19 (1) ◽  
pp. 65-73 ◽  
Author(s):  
Marzieh Ghanemi ◽  
Aminollah Pourshohod ◽  
Mohammad Ali Ghaffari ◽  
Alireza kheirollah ◽  
Mansour Amin ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Head And Neck ◽  
Cell Line ◽  
Squamous Cell ◽  
Optimum Concentration ◽  
Her2 Positive ◽  
Affibody Molecules ◽  
Specific Targeting ◽  
Mcf 7

Background:Expression of human epidermal growth factor receptor type 2 (HER2) in head and neck squamous cell carcinoma (HNSCC) cell line HN5 can be employed with great opportunities of success for specific targeting of anti-cancer chemotherapeutic agents.Objective:In the current study, HER2-specific affibody molecule, ZHER2:342 (an engineered protein with great affinity for HER2 receptors) was selected for conjugation to idarubicin (an anti-neoplastic antibiotic).Method:ZHER2:342 affibody gene with one added cysteine code at the its 5′ end was synthesized de novo and then inserted into pET302 plasmid and transferred to E. Coli BL21 hosting system. After induction of protein expression, the recombinant ZHER2 affibody molecules were purified using Ni- NTA resin and purity was analyzed through SDS-PAGE. Affinity-purified affibody molecules were conjugated to idarubicin through a heterobifunctional crosslinker, sulfosuccinimidyl 4-(Nmaleimidomethyl) cyclohexane-1-carboxylate (Sulfo-SMCC). Specific toxicity of idarubicin-ZHER2 affibody conjugate against two HER2-positive cells, HN5 and MCF-7 was assessed through MTT assay after an exposure time of 48 hours with different concentrations of conjugate.Results:Idarubicin in the non-conjugated form showed potent toxic effects against both cell lines, while HN5 cells were significantly more sensitive compared to MCF-7 cells. Dimeric ZHER2 affibody showed a mild decreasing effect on growth of both HN5 and MCF-7 cells at optimum concentration. Idarubicin-ZHER2 affibody conjugate at an optimum concentration reduced viability of HN5 cell line more efficiently compared to MCF-7 cell line.In conclusion, idarubicin-ZHER2 affibody conjugate in optimum concentrations can be used for specific targeting and killing of HN5 cells.

scholarly journals Comparative Preclinical Evaluation of HER2-Targeting ABD-Fused Affibody® Molecules 177Lu-ABY-271 and 177Lu-ABY-027: Impact of DOTA Position on ABD Domain

Pharmaceutics ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 839
Author(s):  
Yongsheng Liu ◽  
Anzhelika Vorobyeva ◽  
Tianqi Xu ◽  
Anna Orlova ◽  
Annika Loftenius ◽  
...  
Keyword(s):  
A Priori ◽  
Albumin Binding ◽  
Tumor Uptake ◽  
Site Specific ◽  
Affibody Molecules ◽  
Preclinical Evaluation ◽  
Renal Uptake ◽  
C Terminus ◽  
Minimal Modification ◽  
Structure Properties

Radiolabeled Affibody-based targeting agent 177Lu-ABY-027, a fusion of an anti-HER2 Affibody molecule with albumin binding domain (ABD) site-specifically labeled at the C-terminus, has demonstrated a promising biodistribution profile in mice; binding of the construct to albumin prevents glomerular filtration and significantly reduces renal uptake. In this study, we tested the hypothesis that site-specific positioning of the chelator at helix 1 of ABD, at a maximum distance from the albumin binding site, would further increase the strength of binding to albumin and decrease the renal uptake. The new construct, ABY-271 with DOTA conjugated at the back of ABD, has been labelled with 177Lu. Targeting properties of 177Lu-ABY-271 and 177Lu-ABY-027 were compared directly. 177Lu-ABY-271 specifically accumulated in SKOV-3 xenografts in mice. The tumor uptake of 177Lu-ABY-271 exceeded uptake in any other organ 24 h and later after injection. However, the renal uptake of 177Lu-ABY-271 was two-fold higher than the uptake of 177Lu-ABY-027. Thus, the placement of chelator on helix 1 of ABD does not provide desirable reduction of renal uptake. To conclude, minimal modification of the design of Affibody molecules has a strong effect on biodistribution, which cannot be predicted a priori. This necessitates extensive structure-properties relationship studies to find an optimal design of Affibody-based targeting agents for therapy.

scholarly journals Re-targeted adenovirus vectors with dual specificity; binding specificities conferred by two different Affibody molecules in the fiber

Gene Therapy ◽  
2008 ◽  
Vol 16 (2) ◽  
pp. 252-261 ◽  
Author(s):  
S Myhre ◽  
P Henning ◽  
M Friedman ◽  
S Ståhl ◽  
L Lindholm ◽  
...  
Keyword(s):  
Affibody Molecules ◽  
Dual Specificity ◽  
Adenovirus Vectors

scholarly journals Influence of Molecular Design on the Targeting Properties of ABD-Fused Mono- and Bi-Valent Anti-HER3 Affibody Therapeutic Constructs

Cells ◽  
2018 ◽  
Vol 7 (10) ◽  
pp. 164 ◽  
Author(s):  
Mohamed Altai ◽  
Charles Leitao ◽  
Sara Rinne ◽  
Anzhelika Vorobyeva ◽  
Christina Atterby ◽  
...  
Keyword(s):  
Half Life ◽  
Molecular Design ◽  
Growth Factor Receptor ◽  
Fusion Construct ◽  
Affibody Molecules ◽  
Binding Domains ◽  
Biodistribution Studies ◽  
C Terminus ◽  
Type 3

Overexpression of human epidermal growth factor receptor type 3 (HER3) is associated with tumour cell resistance to HER-targeted therapies. Monoclonal antibodies (mAbs) targeting HER3 are currently being investigated for treatment of various types of cancers. Cumulative evidence suggests that affibody molecules may be appropriate alternatives to mAbs. We previously reported a fusion construct (3A3) containing two HER3-targeting affibody molecules flanking an engineered albumin-binding domain (ABD035) included for the extension of half-life in circulation. The 3A3 fusion protein (19.7 kDa) was shown to delay tumour growth in mice bearing HER3-expressing xenografts and was equipotent to the mAb seribantumab. Here, we have designed and explored a series of novel formats of anti-HER3 affibody molecules fused to the ABD in different orientations. All constructs inhibited heregulin-induced phosphorylation in HER3-expressing BxPC-3 and DU-145 cell lines. Biodistribution studies demonstrated extended the half-life of all ABD-fused constructs, although at different levels. The capacity of our ABD-fused proteins to accumulate in HER3-expressing tumours was demonstrated in nude mice bearing BxPC-3 xenografts. Formats where the ABD was located on the C-terminus of affibody binding domains (3A, 33A, and 3A3) provided the best tumour targeting properties in vivo. Further development of these promising candidates for treatment of HER3-overexpressing tumours is therefore justified.

scholarly journals Biodistribution of 211At labeled HER-2 binding affibody molecules in mice

2007 ◽  
Author(s):  
Ann-Charlott Steffen ◽  
Ylva Almqvist ◽  
Ming-Kuan Chyan ◽  
Hans Lundqvist ◽  
Vladimir Tolmachev ◽  
...  
Keyword(s):  
Affibody Molecules ◽  
Her 2

Optimal specific radioactivity of anti-HER2 Affibody molecules enables discrimination between xenografts with high and low HER2 expression levels

2010 ◽  
Vol 38 (3) ◽  
pp. 531-539 ◽  
Author(s):  
Vladimir Tolmachev ◽  
Helena Wållberg ◽  
Mattias Sandström ◽  
Monika Hansson ◽  
Anders Wennborg ◽  
...  
Keyword(s):  
Specific Radioactivity ◽  
Her2 Expression ◽  
Affibody Molecules ◽  
Expression Levels

scholarly journals 17AAG-induced internalisation of HER2-specific Affibody molecules

2016 ◽  
Vol 12 (4) ◽  
pp. 2574-2580 ◽  
Author(s):  
Lovisa Göstring ◽  
Sture Lindegren ◽  
Lars Gedda
Keyword(s):  
Affibody Molecules

scholarly journals HAHAHA, HEHEHE, HIHIHI, or HKHKHK: Influence of Position and Composition of Histidine Containing Tags on Biodistribution of [99mTc(CO)3]+-Labeled Affibody Molecules

2013 ◽  
Vol 56 (12) ◽  
pp. 4966-4974 ◽  
Author(s):  
Camilla Hofström ◽  
Mohamed Altai ◽  
Hadis Honarvar ◽  
Joanna Strand ◽  
Jennie Malmberg ◽  
...  
Keyword(s):  
Affibody Molecules
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