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Multiplex Assay Kit for Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay)

SFTPB; PSPB; SFTB3; SFTP3; SPB; Surfactant Associated Protein B; Pulmonary Surfactant Protein B; 18 kDa pulmonary-surfactant protein; 6 kDa protein

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 79-90 84
EDTA plasma(n=5) 87-99 95
heparin plasma(n=5) 90-98 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 80-97% 80-92% 82-96% 87-94%
EDTA plasma(n=5) 93-104% 89-96% 78-95% 95-104%
heparin plasma(n=5) 93-101% 96-104% 97-104% 82-93%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:SP-B) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Respiratory Physiology & Neurobiology Kinetics of plasma SPB and RAGE during mechanical ventilation in patients undergoing major vascular surgery PubMed: 21736957
American Journal of Rhinology & Allergy Detection of surfactant proteins A, B, C, and D in human nasal mucosa and their regulation in chronic rhinosinusitis with polyps PubMed: 23406594
19 Surfactant protein B and RAGE increases in the plasma during cardiopulmonary bypass: a pilot study PubMed: 20650982
Plos one Staphylococcus aureus and Pseudomonas aeruginosa Express and Secrete Human Surfactant Proteins PubMed: PMC3551896
Plos one The Detection of Surfactant Proteins A, B, C and D in the Human Brain and Their Regulation in Cerebral Infarction, Autoimmune Conditions and Infections of the CNS PubMed: PMC3787032
Cancer Cell. Lung Cancer Signatures in Plasma Based on Proteome Profiling of Mouse Tumor Models PubMed: PMC3406925
Respir Physiol Neurobiol Acute high-altitude exposure reduces lung diffusion: Data from the HIGHCARE Alps project Pubmed: 23619193
PLOS ONE Surfactant-Derived Proteins as Markers of Alveolar Membrane Damage in Heart Failure Pubmed:25514679
Nachweis und Charakterisierung des Oberfl?chenproteins PLUNC (Palate, Lung and Nasal Clone Protein) an der Augenoberfl?che und Bedeutung für das Trockene Auge Opus4:Source
Int J Cardiol Plasma immature form of surfactant protein type B correlates with prognosis in patients with chronic heart failure. A pilot single-center prospective study PubMed: 26310985
Plos One Serum Levels of Surfactant Proteins in Patients with Combined Pulmonary Fibrosis and Emphysema (CPFE) Pubmed:27337142
PLoS One The Cerebral Surfactant System and Its Alteration in HydrocephalicConditions. pubmed:27656877
Frontiers in Aging Neuroscience Correlations of Ventricular Enlargement with Rheologically Active SurfactantProteins in Cerebrospinal Fluid. pubmed:28101052
Frontiers in Aging Neuroscience Correlations of Ventricular Enlargement with Rheologically Active Surfactant Proteins in Cerebrospinal Fluid PMC5209370
Respiratory Physiology & Neurobiology Diving and pulmonary physiology: surfactant binding protein, lung fluid and cardiopulmonary test changes in professional divers pubmed:28467885
Cellular Physiology and Biochemistry Surfactant Protein B Suppresses Lung Cancer Progression by Inhibiting Secretory Phospholipase A2 Activity and Arachidonic Acid Production pubmed:28743125
American Journal of Physiology. Lung Cellular and Molecular Physiology Chronic lung injury and impaired pulmonary function in a mouse model of acid ceramidase deficiency. pubmed:29167126
Respiratory research Effects and molecular mechanisms of intrauterine infection/inflammation on lung development Pubmed:29747649
Respiratory Physiology & Neurobiology  Surfactant proteins changes after acute hemodynamic improvement in patients with advanced chronic heart failure treated with Levosimendan Pubmed:29548887
Lung  Serum Surfactant Protein Levels in Patients Admitted to the Hospital with Acute COPD Exacerbation Pubmed:29445934
American Journal of Physiology Lung Cellular andMolecular Physiology Generation of an alveolar epithelial type II cell line from induced pluripotent stem cells Pubmed: 30211653
Respiratory Research Culture of human alveolar epithelial type II cells by sprouting Pubmed: 30340591
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