Anti-Human folate receptor 1 Recombinant Antibody (Farletuzumab) (CAT#: TAB-113)

Recombinant monoclonal antibody to folate receptor 1. Farletuzumab (MORAb-003) is a monoclonal antibody which is being investigated for the treatment of ovarian cancer, It is targeted at FR-alpha which is overexpressed in some cancers such as ovarian cancer.


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  • COA
PK

Figure 1 The model described the observed plasma farletuzumab concentration data well.

Figure 1 The model described the observed plasma farletuzumab concentration data well.

Farletuzumab final pharmacokinetic model diagnostic plots of observed versus predicted concentrations and conditional residuals versus predicted concentrations.

Farrell, C., Schweizer, C., Wustner, J., Weil, S., Namiki, M., Nakano, T.,... & Phillips, M. D. (2012). Population pharmacokinetics of farletuzumab, a humanized monoclonal antibody against folate receptor alpha, in epithelial ovarian cancer. Cancer chemotherapy and pharmacology, 70(5), 727-734.

PK

Figure 2 The visual predictive check of the final pharmacokinetic model confirmed the suitability of the model.

Figure 2 The visual predictive check of the final pharmacokinetic model confirmed the suitability of the model.

Visual predictive check for the final farletuzumab pharmacokinetic model.

Farrell, C., Schweizer, C., Wustner, J., Weil, S., Namiki, M., Nakano, T.,... & Phillips, M. D. (2012). Population pharmacokinetics of farletuzumab, a humanized monoclonal antibody against folate receptor alpha, in epithelial ovarian cancer. Cancer chemotherapy and pharmacology, 70(5), 727-734.

Cyt

Figure 3 Suppressive effects of sCA125 and mCA125 on farletuzumab-mediated ADCC.

Figure 3 Suppressive effects of sCA125 and mCA125 on farletuzumab-mediated ADCC.

Soluble CA125 inhibited farletuzumab-mediated ADCC against IGROV-1 A. and CAOV-3 B. ovarian cancer cells using primary human PBMCs as effector cells. Shown is percent ADCC-mediated cytotoxicity on target cells at different concentrations of farletuzumab and 30 KU/ml sCA125. * p < 0.05; **p < 0.01. Figures C-E demonstrate the effects of membrane-bound CA125 on ADCC using Jurkat-Luc reporter cells. IGROV-1 C., CAOV-3 D. and OVCAR-3 E. ovarian cancer cell lines were tested for farletuzumab-mediated ADCC activity. The latter two lines were engineered using optimized shRNA vectors to suppress endogenous CA125/MUC16 expression (shown in left graphs where black is CA125 expression in parental cells and red or green in KD cells). Gray peaks represent parental cells and blue peaks represent KD cells stained with secondary antibody only. Farletuzumab-mediated ADCC activation occurred in all parental lines (right-side graphs, blue lines) and was significantly increased in shRNA-CA125 suppressed CAOV-3 (CAOV3-KD red line) and OVCAR-3 (OVCAR-KD1 red line and KD3 green line) cells when incubated with greater than 2.5mg/mL farletuzumab. * p < 0.05; ** p < 0.01.

Kline, J. B., Kennedy, R. P., Albone, E., Chao, Q., Fernando, S., McDonough, J. M.,... & Grasso, L. (2017). Tumor antigen CA125 suppresses antibody-dependent cellular cytotoxicity (ADCC) via direct antibody binding and suppressed Fc-γ receptor engagement. Oncotarget, 8(32), 52045.

Neut

Figure 4 Overview of the BRA assay and effects of sCA125 on CD16a-antibody binding.

Figure 4 Overview of the BRA assay and effects of sCA125 on CD16a-antibody binding.

Microwell plates were coated with farletuzumab, farletuzumab fragments or control agents and Jurkat-CD16a or parental cells analyzed for their ability to adhere to the wall surface. Cells that can adhere remain tethered to the well wall while those that cannot roll and cluster at the U bottom. A. Jurkat cells roll (bottom panel) in contrast to Jurkat-CD16a (top panel) which are retained via farletuzumab Fc-γ receptor binding. B. Wells were coated with farletuzumab or its (Fab') 2 and Jurkat-CD16a are tested for rolling. As shown, cells adhere to the full length farletuzumab (top) but not (Fab') 2 coated wells (bottom) suggesting farletuzumab's Fc domain is required for the Jurkat-CD16a-farletuzumab tethering. C. JurkatCD16a or NK cells were incubated in wells coated with farletuzumab and incubated with or without sCA125. sCA125 inhibited JurkatCD16a-farletuzumab (bottom panel) as well as NK cell-farletuzumab binding (not shown). D. sCA125 was absorbed on CHO-MSLN vs CHO cells and the remaining supernatant was used in ADCC and BRA assays. sCA125-depleted sample buffer no longer inhibited farletuzumab ADCC activity (red bar, top graph) or CD16a-farletuzumab engagement in BRA assay (bottom pictures). CA125-K (blue) is sCA125 absorbed on CHO cells (sCA125 + ); CA125-M (red) is sCA125 absorbed on CHO-MSLN cells (CA125-); CA125-O (green) is starting sCA125 preparation (CA125 + ). E. Full-length farletuzumab and the (Fab') 2 domain are able to compete with sCA125 for JurkatCD16a binding to microwells coated with farletuzumab, suggesting that CA125 perturbs CD16a-farletuzumab binding via interaction within the Fab region. Top row is Jurkat parental. All other rows employ Jurkat-CD16a plus competitors listed on left. Competition assays using: F. neutralizing anti-CD16 antibody; G. excess farletuzumab; or H. sFRA show farletuzumab-CD16a Fc-γ receptor binding specificity for the assay.

Kline, J. B., Kennedy, R. P., Albone, E., Chao, Q., Fernando, S., McDonough, J. M.,... & Grasso, L. (2017). Tumor antigen CA125 suppresses antibody-dependent cellular cytotoxicity (ADCC) via direct antibody binding and suppressed Fc-γ receptor engagement. Oncotarget, 8(32), 52045.

ELISA

Figure 5 ELISA assays using purified reagents demonstrate CA125 binding to farletuzumab and other independent antibodies that result in reduced antibdy-CD16a and-CD32a Fc-γ receptor engagement.

Figure 5 ELISA assays using purified reagents demonstrate CA125 binding to farletuzumab and other independent antibodies that result in reduced antibdy-CD16a and-CD32a Fc-γ receptor engagement.

A. Biotinylated farletuzumab and its (Fab') 2 fragment, but not its Fc fragment bind immobilized sCA125. 96-well plates were coated with 15 KU/mL sCA125 or human serum albumin (HSA) and probed with biotin-labeled farletuzumab, (Fab') 2 or Fc fragments. B. Farletuzumab is able to compete for farletuzumabbiotin binding to sCA125. C. sCA125 suppresses Fc receptor binding to farletuzumab. Farletuzumab was incubated alone or with sCA125 or HSA and probed with biotinylated-CD16a,-CD32a or-CD64a Fc receptor and graphed as a percent inhibition of Fc receptor binding compared to control. sCA125 caused a significant decrease of CD16a binding to farletuzumab as compared to controls (46%, p < 0.0002). Reduction in farletuzumab binding to CD32a Fc receptors was also significant (12%, p < 0.003) while minimal inhibition was observed by sCA125 on farletuzumab binding to CD64a Fc receptor (1%, p = 0.772). Similar reactions were probed with anti-human IgG-HRP to confirm incubation of farletuzumab with sCA125 did not result in less farletuzumab binding to wells (Supplementary Figure 8). D. BRA assay using CHO cells expressing human Fc activating receptors show only farletuzumab-CD16a interactions are disrupted by sCA125. E. Farletuzumab has a significant increase in binding to the mCA125 producing OVCAR parental as compared to mCA125-suppressed isogenic OVCAR-KD1 or mCA125-null CHO membranes. F. sCA125 can bind to subsets of other humanized and fully human mAbs. Seven purified mAbs were biotinylated and used to probe wells coated with 15 KU/mL sCA125 or HSA. While all antibodies were able to robuslty and equally bind their respective target antigen, only a subset were able to specifically bind sCA125 as compared to HSA controls. All ELISAs were done in at least triplicate and different lots of sCA125 or cell membrane preparations were used with similar results. In panels A and B *** p < 0.0001; **** p < 0.00007; ***** p < 0.000001, panels E and F *** p < 0.0001; **** p < 0.00001.

Kline, J. B., Kennedy, R. P., Albone, E., Chao, Q., Fernando, S., McDonough, J. M.,... & Grasso, L. (2017). Tumor antigen CA125 suppresses antibody-dependent cellular cytotoxicity (ADCC) via direct antibody binding and suppressed Fc-γ receptor engagement. Oncotarget, 8(32), 52045.

FC

Figure 6 Effects of sCA125 on ADCC.

Figure 6 Effects of sCA125 on ADCC.

A. FACS analysis of parental CHO and CHO-FRA cells using farletuzumab demonstrates robust cell surface expression of FRA in CHO-FRA cells. Black peak represents unstained cells; blue and green peaks represent farletuzumab-stained CHO and CHO-FRA cells, respectively. B. CHO and CHO-FRA cells were incubated with 10 µg/mL of farletuzumab in the presence of PBMCs. Farletuzumab mediates PBMC ADCC against CHO-FRA but not CHO cells. C. Farletuzumab-mediated NK cell ADCC. CHO and CHO-FRA cells were incubated with 10 μg/mL of farletuzumab in the presence of primary NK cells. sCA125 was found to inhibit farletuzumab-mediated NK cell ADCC in a dose dependent manner. D. sCA125 effects on Jurkat-Luc cells. Jurkat-Luc cells were incubated with 6 µg/mL of farletuzumab and CHO-FRA cells with increasing concentrations of sCA125. As shown, sCA125 inhibits farletuzumab-mediated ADCC signaling in a dose-dependent manner. All data represent values of at least triplicate experiments and all meet p < 0.05.

Kline, J. B., Kennedy, R. P., Albone, E., Chao, Q., Fernando, S., McDonough, J. M.,... & Grasso, L. (2017). Tumor antigen CA125 suppresses antibody-dependent cellular cytotoxicity (ADCC) via direct antibody binding and suppressed Fc-γ receptor engagement. Oncotarget, 8(32), 52045.


Specifications

  • Immunogen
  • Gestational choriocarcinoma cell line, Lu-75.
  • Host Species
  • Human
  • Derivation
  • Humanized (from mouse)
  • Type
  • IgG1 - kappa
  • Specificity
  • Tested positive against native human antigen.
  • Species Reactivity
  • Human
  • Applications
  • FC, IP, ELISA, Neut, FuncS, IF, ICC, PK, Cyt
  • CAS
  • 896723-44-7
  • Generic Name
  • Farletuzumab
  • UNII
  • 2O09BG0OWA
  • MW
  • 145.4 kDa
  • Related Disease
  • Ovarian cancers

Product Property

  • Purity
  • >95.0% as determined by Analysis by RP-HPLC & analysis by SDS-PAGE.
  • Storage
  • Store the antibody (in aliquots) at -20°C. Avoid repeated freezing and thawing of samples.

Applications

  • Application Notes
  • The FOLR1 antibody has been reported in applications of FC, IP, ELISA, Neut, FuncS, IF, ICC, PK, Cyt.

Target

  • Alternative Names
  • Farletuzumab;896723-44-7;MORAb-003;LK26;MORAb-003;FOLR1;folate receptor 1 (adult);FOLR;folate receptor alpha;FR-alpha;KB cells FBP;folate binding protein;folate receptor, adult;adult folate-binding protein;ovarian tumor-associated antigen MOv18;FBP;

Related Resources

  • Biosimilar Overview
Please refer to Farletuzumab Overview to learn more about the mechanism of action, clinical projects, and approved drugs of Farletuzumab.

Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

Downloads

Download resources about recombinant antibody development and antibody engineering to boost your research.

See other products for "Farletuzumab"

Afuco™ Anti-FOLR1 ADCC Recombinant Antibody (Farletuzumab), ADCC Enhanced
This product is an ADCC enhanced antibody produced by our Afuco™ platform. Recombinant monoclonal antibody to folate receptor 1. Farletuzumab (MORAb-003) is a monoclonal antibody which is being investigated for the treatment of ovarian cancer, It is targeted at FR-alpha which is overexpressed in some cancers such as ovarian cancer.

See other products for "FOLR1"

Intrabody

CAT Product Name Application Type
IAB-B029(A) Recombinant Anti-human FOLR1 Intrabody [(D-Arg)9] PCA, Neut, FuncS scFv-(D-Arg)9
IAB-B029(G) Recombinant Anti-human FOLR1 Intrabody [+36 GFP] WB, Neut, FuncS scFv-(+36GFP)
IAB-B029(T) Recombinant Anti-human FOLR1 Intrabody [Tat] WB, ICC, Neut, FuncS scFv-Tat

Chimeric Antibody

CAT Product Name Application Type
TAB-018CQ Human Anti-FOLR1 Recombinant Antibody (TAB-018CQ) ELISA, WB, IHC Chimeric (Rat/Human) IgG1, κ
TAB-018CQ-S(P) Human Anti-FOLR1 Recombinant Antibody; scFv Fragment (TAB-018CQ-S(P)) ELISA, WB, IHC Human scFv
TAB-018CQ-F(E) Human Anti-FOLR1 Recombinant Antibody; Fab Fragment (TAB-018CQ-F(E)) ELISA, WB, IHC Chimeric (Rat/Human) Fab

Low- or Non-fucosylated Oligosaccharide

CAT Product Name Application Type
Gly-029LC-1 Recombinant Anti-Human FOLR1 Antibody (Fc glycosylation/Non fucosylated) ELISA Human antibody

Rabbit Monoclonal Antibody

CAT Product Name Application Type
MOR-1333 Hi-Affi™ Rabbit Anti-FOLR1 Recombinant Antibody (clone DS1333AB) WB Rabbit IgG

ADCC Enhanced Antibody

CAT Product Name Application Type
AFC-TAB-113 Afuco™ Anti-FOLR1 ADCC Recombinant Antibody (Farletuzumab), ADCC Enhanced FC, IP, ELISA, Neut, FuncS, IF ADCC enhanced antibody

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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

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