Publications
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Immunogenicity of primary and booster MenACWY-TT vaccination in older adults and the importance of IgM
Where the paper cites Pel-Freez
…Frailty Indicator self-report questionnaire35. Serological analysis Functional MenACWY-specific antibody titers were determined using the serum bactericidal assay with baby rabbit complement (rSBA) (Pelfreez, lot: 21044-C) as described previously15,36. Target strains included MenA (strain 3125), MenC (strain C11), MenW (strain MP01240070), and MenY (strain S-1975). rSBA titers were expressed as the reciprocal…
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Divergent humoral immune signatures in response to Klebsiella pneumoniae bacteremia
Where the paper cites Pel-Freez
…combined with 20 µl of heat-inactivated human serum (inactivated at 56°C for 30 minutes) in 96-well U-bottom plates and incubated shaking at 37°C for 1 h. Subsequently, 10 µl of baby rabbit complement (Pel-Freez Biologics) was added to a final concentration of 10%, and plates were incubated shaking for 1 h at 37°C. All samples were tested in triplicate. Controls included bacteria alone, bacteria with serum only,…
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Naturally acquired antibodies against 4 Streptococcus pneumoniae serotypes in Pakistani adults with type 2 diabetes mellitus
Where the paper cites Pel-Freez
…strains of pneumococcal serotype 19F, 18C, and 9V. Plates were incubated for 30 minutes on a shaker (750 RPM) at room temperature, followed by the addition of 10 μL baby rabbit complement (Pel-freeze biologicals) and differentiated HL-60 cells (cells: bacteria was 200:1). Plates were incubated for another 45 minutes at 37°C in a humidified chamber, followed by cooling on ice for 15 minutes to stop…
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Three neutralizing mAbs induced by MPXV A29L protein recognizing different epitopes act synergistically against orthopoxvirus
Where the paper cites Pel-Freez
…assay The neutralizing activity of the mAbs and sera was determined using the MV or EV forms of the VACV Tian tan or WR strain. Neutralization of VACV EV was performed using a 10% baby rabbit complement (Pelfreez). For EV neutralization, MV was depleted with blocking mAbs at 20 mg/mL. BHK-21 cells were maintained in DMEM sup- plemented with 10% calf serum at 37°C, and 5% CO2. On the day of the experiment, BHK-21…
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The Group A Streptococcal Vaccine Candidate VAX-A1 Protects against Group B Streptococcus Infection via Cross-Reactive IgG Targeting Virulence Factor C5a Peptidase
Where the paper cites Pel-Freez
…to the manufacturer’s protocol. Neutrophils were adjusted to a concentration of 5 × 10 6 cells/mL in RPMI 1640 tissue culture media, and the bacteria were pre-incubated with baby rabbit complement (PelFreez, #31061) and heat-inactivated fetal bovine serum (FBS, Cat# 97068-085, VWR International). GBS strains were grown to the mid-logarithmic growth phase of OD 600 = ~0.4 and washed twice with sterile…
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Gamma-Irradiated Non-Capsule Group B Streptococcus Promotes T-Cell Dependent Immunity and Provides a Cross-Protective Reaction
Where the paper cites Pel-Freez
…adjusted to 10 5 CFU/mL. Following incubation at room temperature for 30 min with shaking, 40 µL of a differentiated HL-60 cell suspension containing 4 × 10 5 cells and 10 µL baby rabbit complement (BRC; Pel-Freez Biological: Rogers AR) was added. The cell mixture was incubated for 45 min at 37 °C with shaking. Next, 10 µL of the final mixture was spotted onto THY agar containing 1.5% Bacto agar (BD Biosciences).…
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The Lack of Natural IgM Increases Susceptibility and Impairs Anti-Vi Polysaccharide IgG Responses in a Mouse Model of Typhoid
Where the paper cites Pel-Freez
…Ty2 in DPBS (250–500 CFU) was added to each well of a round-bottom polypropylene 96-well plate containing 50 μl of heat-inactivated serum in serial dilutions, 12.5 μl of baby rabbit complement (Pel-Freez Biologicals, Rogers, AR), and 27.5 μl of DPBS. Triplicate samples of each dilution were incubated for 120 min at 37°C with gentle rocking, and 10 μl of this mixture were plated on LB agar plates for counting…
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Performance of a Four-Antigen Staphylococcus aureus Vaccine in Preclinical Models of Invasive S. aureus Disease
Where the paper cites Pel-Freez
…immune sera were mixed with either a CP5-expressing or CP8-expressing clinical isolate of S. aureus and allowed to opsonize the bacteria. The reaction mixtures were then mixed with baby rabbit complement (Pel-Freez, Rogers, AR, USA) and neutrophil-like HL-60 cells (ATCC ® CCL-240™, Manassas, VA, USA). An OPA antibody titer was defined as the reciprocal of the highest serum dilution resulting in 50% reduction of the…
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Multicomponent Vaccines against Group A Streptococcus Can Effectively Target Broad Disease Presentations
Where the paper cites Pel-Freez
…penicillin–streptomycin and differentiated with 0.8% DMF at 4 × 10 5 cells/mL in 100 mL for 5 to 6 days. Frozen GAS cultures were tested with differentiated HL60 cells and 2% baby rabbit complement (Pel-Freez Biologicals) or heat inactivated complement at a bacteria:cell ratio of 1:400 with <35% nonspecific killing [ 13 ]. To perform the OPA, GAS frozen cultures were thawed, washed and diluted to the required input…
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Evaluation of Critical Quality Attributes of a Pentavalent (A, C, Y, W, X) Meningococcal Conjugate Vaccine for Global Use
Where the paper cites Pel-Freez
…Pathogens 2021, 10, 928 15 of 17 To determine rSBA titers, individual rabbit sera were inactivated at 56 ◦C before initiating the assay. The complement source was 3–4 week old rabbit complement (Pel- freez Biologicals, Rogers, AR, USA). The bacterial strains used in the rSBA assays were: serogroup A, CDC F8238, phenotype A:4:P1.9, O-Acetylation OAc+; serogroup C, strain C11, C:16:P1.7–1,1.1, OAc+; serogroup Y,…
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Development and Immunogenicity of a Prototype Multivalent Group B Streptococcus Bioconjugate Vaccine
Where the paper cites Pel-Freez
…Bio-Rad). The samples were subsequently transferred to nitrocellulose membranes with a 0.2 μm pore size (Bio-Rad… Baby rabbit complement (Pel-Freez) was added to HL-60 cells at a 1…
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Development and Immunogenicity of a Brazilian Glycoconjugate vaccine against Meningococcal W in a Pilot Scale
Where the paper cites Pel-Freez
…Pel-Freez® baby rabbit complement lacking bactericidal activity was added. After incubation 10 µl of the mixture from each well was plated out on Columbia blood agar plate by the tilt…
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Broadly Reactive Human Monoclonal Antibodies Targeting the Pneumococcal Histidine Triad Protein Protect against Fatal Pneumococcal Infection
Where the paper cites Pel-Freez
…were mixed with HBSS as a negative control, and purified human serum IgG was used as a positive control. Differentiated HL-60 cells were washed twice with HBSS and mixed with baby rabbit complement (Pel-Freez Biologicals) at a final concentration of 10% in each well. Following this, 60ml (1 106 viable cells) of differentiated HL60 cells and complement were added to the mixture of bacteria and antibodies and…
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A Structural Model for the Ligand Binding of Pneumococcal Serotype 3 Capsular Polysaccharide-Specific Protective Antibodies.
Where the paper cites Pel-Freez
…and 1% L-glutamine. HL-60 cells were differentiated using 0.6% N,N-dimethylformamide (DMF) for 3 days before performing the OPKA assay, harvested, and resuspended in OBB. Baby rabbit complement (Pel- Freez) was added to HL-60 cells at a 1:5 final volume. The HL-60-complement mixture was added to the bacteria at 5 105 cells/well. The final reaction mixtures were incubated at 37°C for 1 h with shaking. The…
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Age-stratified analysis of serotype-specific baseline immunity against group B streptococcus
Where the paper cites Pel-Freez
…plates. After 30 min of Accepted Manuscript 7 incubation at room temperature, 42.8 μl of a differentiated HL-60 cell suspension (4.28 × 105 cells per well) and 7.2 μl of baby rabbit complement (BRC; Pel-Freez Biological; Rogers, AR) were added to each well, after which the mixture was incubated for 45 min at 37 °C with shaking. Afterwards, 10 µL of the final reaction mixture from each well was spotted onto…
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T Cell–Mediated Humoral Immune Responses to Type 3 Capsular Polysaccharide of Streptococcus pneumoniae
Where the paper cites Pel-Freez
…were cultured in RPMI 1640 with 10% heat- inactivated FetalClone (SH30080.03; HyClone) and 1% L-glutamine. HL60 cells were differentiated as previously described (15). Baby rabbit comple- ment (31061; Pel-Freez) was added to HL60 cells at 20% final volume, with a final cell concentration of 1 3 107 cells/ml. The HL60/complement mix- ture was added to the serum/bacteria at 5 3 105 cells per well. The reactions…
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Functional Assays for B Cells and Antibodies
Where the paper cites Pel-Freez
…HL-60 cell maintenance) (no. SH30080.03; HyClone) Premium fetal bovine serum (FBS, for preparation of assay buffer [OBB]) (no. 11150; Atlanta Biologicals) Baby rabbit complement (BRC) (no. 31061; Pel-Freez Biologicals) HL-60 cell line (no. CCL240; ATCC) Blood agar plates Glycerol Gelatin Dimethyl sulfoxide (DMSO) Target bacteria (see Table 4) Prepared Solutions Streptomycin stock solution Prepare a 300 mg/ml…
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Functional antibodies to Haemophilus influenzae type B, Neisseria meningitidis, and Streptococcus pneumoniae contained in intravenous immunoglobulin products
Where the paper cites Pel-Freez
…salt solution (HBSS) with Ca21 and Mg21 (Life Technologies) and 0.1% gelatin and aliquoted in a 96-well microplate. Diluted Hib Eagan strain suspension (10 mL) and 10 mL of baby rabbit complement (Pel-Freeze Biologicals) were added in the 96-well microplate. The mixture was incu- bated for 2 minutes at 258C with shaking at 700 rpm and then incubated for 30 min at 378C/5% CO2 without shak- ing. After stopping the…
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Functional immune responses to twelve serotypes after immunization with a 23-valent pneumococcal polysaccharide vaccine in older adults
Where the paper cites Pel-Freez
…CFU in each well of a 96-well microtiter plate. After 30 min of incubation at room temperature with shaking, 40 L of HL-60 cell suspension (4 × 105 cells per well) and 10 L of baby rabbit complement (Pel-Freez, Brown Deer, WI, USA) were added to each well. Plates were incubated in a tissue culture incubator (37 ◦C, 5% CO2) with shaking for 45 min. An aliquot of the final reac- tion mixture (10 L) was spotted…
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Impact of immunization with Protein F on pulmonary clearance of nontypeable Haemophilus influenzae
Where the paper cites Pel-Freez
…of rabbit anti-PF Abs. Briefly, differentiation of HL60 cells into granulocytes was induced by suspending cells into culture medium containing 1.25% dimethyl sulfoxide for 5 days. Baby rabbit complement (Pel-freez Biologi- cals Rogers, AR) was used as a standardized source of complement. Bacteria were pre-incubated with various titers of affinity-purified Abs for 30 min at RT on a mini-orbital shaker. Subsequently,…
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Myeloablative therapy with autologous bone marrow transplantation as consolidation of remission in patients with follicular lymphoma
Where the paper cites Pel-Freez
…non-Hodgkin’s lymphoma. The marrow mononuclear cell fraction is being treated in vitro with three cycles of the monoclonal antibody anti-CD20 (anti-B1, Coulter Immunology) and baby rabbit complement (Pel-Freez). Thirty-eight patients with follicular lymphoma (age range 29–61 years, median 43) have been treated to date. At the time of treatment, 28 patients were in second remission, 7 were in third, and 3 were…