A novel method for measuring cellular antibody uptake using imaging flow cytometry reveals distinct uptake rates for two different monoclonal antibodies targeting L1
Monoclonal antibodies (mAbs) have emerged as a promising tool for cancer therapy. Differing approaches utilize mAbs to either deliver a drug to the tumor cells or to modulate the host's immune system to mediate tumor kill. The rate by which a therapeutic antibody is being internalized by tumor...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article (Journal) |
| Language: | English |
| Published: |
9 May 2015
|
| In: |
Journal of immunological methods
Year: 2015, Volume: 423, Pages: 70-77 |
| ISSN: | 1872-7905 |
| DOI: | 10.1016/j.jim.2015.04.024 |
| Online Access: | Verlag, Volltext: https://doi.org/10.1016/j.jim.2015.04.024 Verlag: http://www.sciencedirect.com/science/article/pii/S0022175915001519 
|
| Author Notes: | John Hazin, Gerhard Moldenhauer, Peter Altevogt, Nathan R. Brady |
MARC
| LEADER | 00000caa a2200000 c 4500 | ||
|---|---|---|---|
| 001 | 1724943332 | ||
| 003 | DE-627 | ||
| 005 | 20230426103314.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 200716s2015 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1016/j.jim.2015.04.024 |2 doi | |
| 035 | |a (DE-627)1724943332 | ||
| 035 | |a (DE-599)KXP1724943332 | ||
| 035 | |a (OCoLC)1341345607 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rda | ||
| 041 | |a eng | ||
| 084 | |a 33 |2 sdnb | ||
| 100 | 1 | |a Hazin, John |e VerfasserIn |0 (DE-588)1071579789 |0 (DE-627)826057292 |0 (DE-576)43316607X |4 aut | |
| 245 | 1 | 2 | |a A novel method for measuring cellular antibody uptake using imaging flow cytometry reveals distinct uptake rates for two different monoclonal antibodies targeting L1 |c John Hazin, Gerhard Moldenhauer, Peter Altevogt, Nathan R. Brady |
| 264 | 1 | |c 9 May 2015 | |
| 300 | |a 8 | ||
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Gesehen am 16.07.2020 | ||
| 520 | |a Monoclonal antibodies (mAbs) have emerged as a promising tool for cancer therapy. Differing approaches utilize mAbs to either deliver a drug to the tumor cells or to modulate the host's immune system to mediate tumor kill. The rate by which a therapeutic antibody is being internalized by tumor cells is a decisive feature for choosing the appropriate treatment strategy. We herein present a novel method to effectively quantitate antibody uptake of tumor cells by using image-based flow cytometry, which combines image analysis with high throughput of sample numbers and sample size. The use of this method is established by determining uptake rate of an anti-EpCAM antibody (HEA125), from single cell measurements of plasma membrane versus internalized antibody, in conjunction with inhibitors of endocytosis. The method is then applied to two mAbs (L1-9.3, L1-OV52.24) targeting the neural cell adhesion molecule L1 (L1CAM) at two different epitopes. Based on median cell population responses, we find that mAb L1-OV52.24 is rapidly internalized by the ovarian carcinoma cell line SKOV3ip while L1 mAb 9.3 is mainly retained at the cell surface. These findings suggest the L1 mAb OV52.24 as a candidate to be further developed for drug-delivery to cancer cells, while L1-9.3 may be optimized to tag the tumor cells and stimulate immunogenic cancer cell killing. Furthermore, when analyzing cell-to-cell variability, we observed L1 mAb OV52.24 rapidly transition into a subpopulation with high-internalization capacity. In summary, this novel high-content method for measuring antibody internalization rate provides a high level of accuracy and sensitivity for cell population measurements and reveals further biologically relevant information when taking into account cellular heterogeneity. | ||
| 650 | 4 | |a Antibody internalization | |
| 650 | 4 | |a Endocytosis | |
| 650 | 4 | |a EpCAM | |
| 650 | 4 | |a Imaging flow cytometry | |
| 650 | 4 | |a L1 | |
| 650 | 4 | |a L1CAM | |
| 700 | 1 | |a Moldenhauer, Gerhard |e VerfasserIn |0 (DE-588)1054128634 |0 (DE-627)791055701 |0 (DE-576)410008443 |4 aut | |
| 700 | 1 | |a Altevogt, Peter |d 1947- |e VerfasserIn |0 (DE-588)1113206306 |0 (DE-627)867118784 |0 (DE-576)164729240 |4 aut | |
| 700 | 1 | |a Brady, Nathan |e VerfasserIn |0 (DE-588)1075765625 |0 (DE-627)833820230 |0 (DE-576)444637591 |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Journal of immunological methods |d Amsterdam [u.a.] : Elsevier Science, 1971 |g 423(2015), Seite 70-77 |h Online-Ressource |w (DE-627)306659751 |w (DE-600)1500495-8 |w (DE-576)081986386 |x 1872-7905 |7 nnas |a A novel method for measuring cellular antibody uptake using imaging flow cytometry reveals distinct uptake rates for two different monoclonal antibodies targeting L1 |
| 773 | 1 | 8 | |g volume:423 |g year:2015 |g pages:70-77 |g extent:8 |a A novel method for measuring cellular antibody uptake using imaging flow cytometry reveals distinct uptake rates for two different monoclonal antibodies targeting L1 |
| 856 | 4 | 0 | |u https://doi.org/10.1016/j.jim.2015.04.024 |x Verlag |x Resolving-System |3 Volltext |
| 856 | 4 | 0 | |u http://www.sciencedirect.com/science/article/pii/S0022175915001519 |x Verlag |
| 951 | |a AR | ||
| 992 | |a 20200716 | ||
| 993 | |a Article | ||
| 994 | |a 2015 | ||
| 998 | |g 1075765625 |a Brady, Nathan |m 1075765625:Brady, Nathan |p 4 |y j | ||
| 998 | |g 1113206306 |a Altevogt, Peter |m 1113206306:Altevogt, Peter |d 60000 |d 61900 |d 50000 |e 60000PA1113206306 |e 61900PA1113206306 |e 50000PA1113206306 |k 0/60000/ |k 1/60000/61900/ |k 0/50000/ |p 3 | ||
| 998 | |g 1054128634 |a Moldenhauer, Gerhard |m 1054128634:Moldenhauer, Gerhard |d 910000 |e 910000PM1054128634 |k 0/910000/ |p 2 | ||
| 998 | |g 1071579789 |a Hazin, John |m 1071579789:Hazin, John |p 1 |x j | ||
| 999 | |a KXP-PPN1724943332 |e 3724761287 | ||
| BIB | |a Y | ||
| SER | |a journal | ||
| JSO | |a {"physDesc":[{"extent":"8 S."}],"relHost":[{"pubHistory":["1.1971/72 -"],"titleAlt":[{"title":"JIM"}],"part":{"text":"423(2015), Seite 70-77","volume":"423","extent":"8","year":"2015","pages":"70-77"},"note":["Gesehen am 30.01.20"],"disp":"A novel method for measuring cellular antibody uptake using imaging flow cytometry reveals distinct uptake rates for two different monoclonal antibodies targeting L1Journal of immunological methods","type":{"media":"Online-Ressource","bibl":"periodical"},"recId":"306659751","language":["eng"],"title":[{"title_sort":"Journal of immunological methods","title":"Journal of immunological methods","subtitle":"JIM"}],"physDesc":[{"extent":"Online-Ressource"}],"origin":[{"dateIssuedKey":"1971","publisher":"Elsevier Science","dateIssuedDisp":"1971-","publisherPlace":"Amsterdam [u.a.]"}],"id":{"issn":["1872-7905"],"eki":["306659751"],"zdb":["1500495-8"]}}],"origin":[{"dateIssuedDisp":"9 May 2015","dateIssuedKey":"2015"}],"id":{"eki":["1724943332"],"doi":["10.1016/j.jim.2015.04.024"]},"name":{"displayForm":["John Hazin, Gerhard Moldenhauer, Peter Altevogt, Nathan R. Brady"]},"type":{"media":"Online-Ressource","bibl":"article-journal"},"note":["Gesehen am 16.07.2020"],"language":["eng"],"recId":"1724943332","title":[{"title_sort":"novel method for measuring cellular antibody uptake using imaging flow cytometry reveals distinct uptake rates for two different monoclonal antibodies targeting L1","title":"A novel method for measuring cellular antibody uptake using imaging flow cytometry reveals distinct uptake rates for two different monoclonal antibodies targeting L1"}],"person":[{"roleDisplay":"VerfasserIn","display":"Hazin, John","role":"aut","family":"Hazin","given":"John"},{"roleDisplay":"VerfasserIn","display":"Moldenhauer, Gerhard","role":"aut","family":"Moldenhauer","given":"Gerhard"},{"roleDisplay":"VerfasserIn","display":"Altevogt, Peter","role":"aut","family":"Altevogt","given":"Peter"},{"display":"Brady, Nathan","roleDisplay":"VerfasserIn","role":"aut","family":"Brady","given":"Nathan"}]} | ||
| SRT | |a HAZINJOHNMNOVELMETHO9201 | ||