Measurement of mouse anti-phospholipid antibodies to solid-phase microspheres by both flow cytofluorometry and Alcian blue-pretreated microtitre plates in an ELISA

Laakel, Mohamed; Bouchard, Martin; Lagacé, Josée

doi:10.1016/0022-1759(95)00282-0

Cited by 15 publications

(5 citation statements)

References 11 publications

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“…This manifests as a surprisingly robust, long-lived humoral immune response to PEG that is sufficient to cause accelerated blood clearance and loss of disease site targeting upon subsequent re-administration. Liposomes incorporating immunostimulatory molecules are known to act as potent adjuvants that can promote Ab responses against weakly immunogenic antigens [19,20], including lipids [27,28], displayed on the outer surface of the liposome. Reports have also shown that Ab responses against PEG can be raised when PEGylated proteins are used in conjunction with aggressive immunization regimens [29,30].…”

Section: Disease Site Targeting and Blood Clearance Of Pegylated Lipomentioning

confidence: 99%

Hypersensitivity and Loss of Disease Site Targeting Caused by Antibody Responses to PEGylated Liposomes

Judge¹,

McClintock²,

Phelps³

et al. 2006

Molecular Therapy

241

164

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The systemic application of nucleic acid drugs requires delivery systems that overcome the poor pharmacokinetics, limited biodistribution, and inefficient uptake of nucleic acids. PEGylated liposomes show considerable promise because of their intrinsic ability to accumulate at disease sites and facilitate transfection of target cells. Unlike many viral vectors, PEGylated liposomes are generally considered to be nonimmunogenic. We have developed a PEGylated liposome for the systemic administration of plasmid DNA that achieves high levels of selective gene expression at distal tumor sites. Here we report that the in vivo efficacy and safety of these systems can be severely compromised following repeat administration. This phenomenon is characterized by a loss of disease site targeting, accelerated clearance from the blood, and acute hypersensitivity. These effects are fully attributable to a surprisingly robust, long-lived antibody response generated against polyethylene glycol (PEG) that results from the strong adjuvant effect of the plasmid payload. Importantly, immunogenicity may be substantially reduced by modifying the alkyl chain of the PEG-lipid conjugate, thereby allowing successful repeat dosing of the modified plasmid formulations without adverse side effects. Immunogenicity is a relevant concern for a number of nonviral delivery systems given the potent immunostimulatory properties of many nucleic acid drugs.

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Section: Disease Site Targeting and Blood Clearance Of Pegylated Lipomentioning

confidence: 99%

Hypersensitivity and Loss of Disease Site Targeting Caused by Antibody Responses to PEGylated Liposomes

Judge¹,

McClintock²,

Phelps³

et al. 2006

Molecular Therapy

241

164

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show abstract

“…Beads coated with phosphatidylinositol (PI) or PS were also tested although the frequency of samples positive by flow cytometry and ELISA were not compared. This methodology has recently been shown to be reproducible and sensitive for the detection of mouse anti-phospholipid antibodies (26). The apolipoprotein ß 2 glycoprotein 1 (ß 2 gp1) has been shown to be necessary for some anti-CL antibodies to bind in ELISA (22)(23)(24) and a similar observation has been reported using flow cytometry (21).…”

mentioning

confidence: 83%

Detection of anti‐phosphatidylethanolamine antibodies using flow cytometry

Drouvalakis¹,

Neeson²,

Buchanan³

1999

Cytometry

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Section: Introductionmentioning

confidence: 99%

“…This in turn allows us to analyze higher volumes of samples as we can reduce the possibility of background noise while increasing the likelihood of capturing a greater number of the target of interest as well the use of a capture antibody can decrease risks of cross-reactive detection. 22,23 Here, we present a solid phase-based variant of the proximity extension assay (SP-PEA), which serves to improve the efficiency of detection to improve detection sensitivity and dynamic range of detection compared to homogenous PEA (Figure 1.). Target molecules are first captured from liquid samples via immobilized antibodies on magnetic beads.…”

Section: Introductionmentioning

confidence: 99%

Magnetic Bead Based Proximity Extension Assay for Sensitive Protein and Extracellular Vesicles Detection

Muthelo

Ebai

Doulabi

et al. 2022

Preprint

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During the early stages of disease development, protein biomarkers can leak into the blood creating opportunities for early diagnosis of disease with minimally invasive sampling. These proteins biomarkers however are often masked by the presence of more abundant functional blood proteins, making specific detection a challenge with most current immunoassays. We we report on the development of a magnetic bead based solid-phase PEA (SP-PEA) for sensitive detection of proteins in plasma and serum samples. Antibody functionalized magnetic beads are used to capture the target of interest. Following capture, non-specifically bound proteins are washed off before PEA probes are added for detection of the bound proteins. Compared to hoogenous PEA, SP-PEA admits the use of larger sample volumes to increase available target molecules, higher concentration of detection reagents for more efficient formation of detection complexes and washes for removal of nonspecific background. We compared SP-PEA to solution phase PEA for the detection of cytokines: interlukin-6, interlukin-2, interlukin-4, interlukin-10 and Tumor Necrosis Factor-alpha, and we demonstrated an increased sensitivity by 15 to 60 fold in buffer and chicken serum. We further expanded SP-PEA to detect extracellular vesicles (EVs) through combinations of proteins on the surface of specific EV populations.

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Measurement of mouse anti-phospholipid antibodies to solid-phase microspheres by both flow cytofluorometry and Alcian blue-pretreated microtitre plates in an ELISA

Cited by 15 publications

References 11 publications

Hypersensitivity and Loss of Disease Site Targeting Caused by Antibody Responses to PEGylated Liposomes

Hypersensitivity and Loss of Disease Site Targeting Caused by Antibody Responses to PEGylated Liposomes

Detection of anti‐phosphatidylethanolamine antibodies using flow cytometry

Magnetic Bead Based Proximity Extension Assay for Sensitive Protein and Extracellular Vesicles Detection

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