Antibiotic and Antibacterial Vaccine Development Services

06/01/18

www.SouthernResearch.org/Drug-Development Contents

Why choose Southern Research? ...... 4 in vitro Testing Services...... 5 B0101 Minimum Inhibitory Concentration (MIC): ...... 5 B0102 Minimum Bactericidal Concentration (MBC): ...... 5 B0103 Time-kill Assay: ...... 5 B0104 Resistance Frequency Determination: ...... 5 B0105 Post-antibiotic Effect (PAE): ...... 6 B0106 Serum and B0107 Protein Sensitivity: ...... 6 B0108 Serum and B0109 Protein Binding: ...... 6 B0110 Plasma and B0111 Microsome Stability: ...... 6 B0112 Synergy Testing: ...... 7 B0113 Minimum Biofilm Inhibition Concentration (MBIC): ...... 7 B0114 Minimum Biofilm Eradication Concentration (MBEC): ...... 7 B0115 Minimum Inhibitory Concentration 90% (MIC 90): ...... 8 B0116 Opsonophagocytic Killing Assay (OPKA): ...... 8 B0117 Opsonophagocytic Uptake Assay (OPUA): ...... 8 B0118 Serum Bactericidal Activity (SBA): ...... 8 in vivo Testing ...... 9 B0201 Maximum tolerated dose (MTD): ...... 9 B0202 Dose Range Finding (DRF): ...... 9 B0203 Pharmacokinetic Profiling (PK): ...... 9 B0204 Bioavailability: ...... 9 B0205 Neutropenic Mouse Thigh (NMTh): ...... 10 B0206 Neutropenic Mouse Lung (NMLu): ...... 10 B0207 Acute Murine Sepsis/Peritonitis: ...... 10 B0208 Chronic Murine Sepsis (Cecal Ligation and Puncture): ...... 10 B0209 Murine Acute Pneumonia: ...... 11 B0210 Skin Infection – Tape-stripping: ...... 11 B0211 Surgical Site Infection—Suture: ...... 11

www.SouthernResearch.org/Drug-Development B0212 Wound Infection—Punch Biopsy: ...... 11 B0213 Transcervical Chlamydia Persistence: ...... 12 B0214 Pharmacodynamic Dose Fractionation: ...... 12 B0215 in vivo Resistance Monitoring: ...... 12 B0216 in vivo Post-Antibiotic Effect (PAE): ...... 12 B0217 Vaccine Immunogenicity Evaluation: ...... 13 IND-Enabling Toxicology: ...... 13

Clinical Trial Endpoint Analysis ...... 14 B0301 Opsonophagocytic Killing Assay (OPKA): ...... 14 B0302 Opsonophagocytic Uptake Assay (OPUA): ...... 14 B0303 Serum Bactericidal Activity (SBA): ...... 14 Additional Bioanalytical and Immunological Endpoint Assays: ...... 14

www.SouthernResearch.org/Drug-Development Southern Research Why choose Southern Research? Founded in 1941 in Birmingham, Alabama, Southern Research is a scientific and engineering contract research organization that conducts drug discovery and development, environmental and occupational hazard characterization, advanced engineering research in materials and systems development, and energy and environmental technologies research. SR supports clients and partners in the pharmaceutical, agrochemical, biotechnology, defense, aerospace, environmental, and energy industries. We deliver quality results and a quality experience.

That means we’re rapid, reliable, and responsive. With over 50 years of experience, we recognized that no two test articles are ever the same. That’s why our approach has always been, and still is, to focus on each client’s individual needs. Our scientists and study directors will take the time to understand your program and the specific risks and liabilities you are concerned about. Then, after we fully understand your needs, we will work with you to develop the most appropriate, regulatory-compliant testing plan.

But the quality experience doesn’t stop when the study protocol is signed. Our nationally recognized scientific and quality assurance teams will execute the plan to our exacting operational standards, ensuring we generate the highest quality data and reports. Throughout the study, be it one day or two years, your Southern Research study director will be readily accessible and will proactively communicate the status of your project. Services What our clients are saying . In vitro antibacterial, antiviral and anticancer screening . “Everything has been excellent. Communication between . Infectious disease and cancer animal efficacy models my company and the SR group has been great. They have been very responsive and have expertly solved all issues . Pharmacokinetics/Toxicokinetics we’ve encountered. The SR group has performed . Immunoassay and bioanalytical chemistry exceptionally well! We hope to work with them again.” . Immunogenicity and immunopotency . “I’m very impressed with the SR study director. He is very . Vaccine neurovirulence/neuropathology responsive and thoughtful. I very much enjoy working with . IND-enabling nonclinical toxicology him.” . . Developmental and reproductive toxicology “Thanks for all of your efforts. The study, although very challenging, really went a long way towards us obtaining . Neurotoxicity and neurobehavioral assessment clearance to commence our Phase I trial.” . Clinical trial endpoint analysis

Experience The SR campus in downtown Birmingham, Alabama. . Internationally recognized infectious disease, cancer, immunoassay, and bioanalytical chemistry experts . Multiple American Board of Toxicology certified toxicologists . Multiple American College of Veterinary Pathologists certified pathologists . Multi-award winning histopathology support team . American Association for Laboratory Animal Science (AALAS) accredited technicians and managers . Nationally recognized quality assurance unit . Compliance with federal and industry regulations (ICH, OECD, EPA, FDA, USDA, OLAW, GLP)

www.SouthernResearch.org/Drug-Development Southern Research | in vitro Services in vitro Testing Services

B0101 Minimum Inhibitory Concentration (MIC): The MIC is defined as the lowest concentration of compound needed to inhibit visible growth for a given organism. It is determined following Clinical and Laboratory Standards Institute (CLSI) guidelines, as appropriate. The MIC is reported as the lowest concentration of drug that completely inhibits growth of the test organism. Southern Research has pre-assembled panels of test organisms to ease strain selection including: An ESKAPE pathogen panel, an MDR panel, a CRE panel, an ESBL-panel, a respiratory pathogen panel, a wound pathogen panel, a CF pathogen panel, an STI pathogen panel, a meningitis panel, an endocarditis panel, and a gastroenteritis panel.

Reportable Results: Compound concentration (µg/mL)

B0102 Minimum Bactericidal Concentration (MBC):

The MBC is defined as the lowest concentration of compound that achieves at least a 3 log10 reduction in CFU compared to the initial inoculum. It is determined subsequent to MIC testing by subculturing diluted aliquots to a solid medium for bacterial enumeration. These aliquots are taken from wells containing compound but that fail to exhibit macroscopic growth. MBC values greater than 16 times the MIC typically indicate bacteriostatic activity or antimicrobial tolerance.

Reportable Results: Compound concentration (µg/mL)

B0103 Time-kill Assay: The time-kill or Suspension Test determines the time required for the antimicrobial agent to inactivate the challenge test microorganism. Microbial kill is measured as a function of time and concentration. Typically, testing is conducted over a 24-hour time period using concentrations of compound at 1-2X the established MIC. Aliquots are plated at various time-points to determine bacterial viability.

Reportable Results: Log10 CFU v. Time

B0104 Resistance Frequency Determination: To determine the spontaneous resistance frequency, bacteria are grown to late Log phase, washed, and plated onto agar plates containing the test article at 4 x the MIC. To determine the induced resistance frequency, bacteria are grown to late log phase or stationary phase in sub lethal concentrations of the test article before washing and plating. In both cases, the number of resistant colonies is divided by the total viable cell count per plate to calculate the resistance frequency.

Reportable Results: Frequency of Resistance (resistant colonies/total viable cell count)

www.SouthernResearch.org/Drug-Development Southern Research | in vitro Services B0105 Post-antibiotic Effect (PAE): The post-antibiotic effect (PAE) is an important pharmacodynamics (PD) parameter that evaluates the ability of a given test article to suppress bacterial growth after drug levels fall below the MIC. Bacteria in the log phase of growth are exposed to 2X, 4X and 8X the MIC for 1 hour. After exposure, cells are washed to remove the test article and suspended in broth without antibiotic. Viable bacteria are enumerated by serial plating at hourly time points for 6-8 hours.

Reportable Results: PAE is the time (in hours) for drug exposed bacteria to increase in number by 1

log10 minus the time for non-exposed cells to increase in number by 1 log10.

B0106 Serum and B0107 Protein Sensitivity: The pharmacodynamics (PD) of antibiotics can be substantially altered due to binding to protein in serum. To screen for this possibility, time-kill assays are performed at 2X, 5X, 10X, 20X and 50X the MIC in media supplemented with 20 g/L human albumin (the albumin content of interstitial fluid) or inactivated serum (50% v/v).

Reportable Results: Log10 CFU v. Time for each condition in standard media and media supplemented with albumin or serum.

B0108 Serum and B0109 Protein Binding: Protein binding is determined by a standard dialysis method wherein the test article is suspended in a solution containing active or inactivated human serum 50% v/v (B0108 Serum Binding) or 20 g/L or 40 g/L albumin (B0109 Protein Binding). During dialysis, the solution is allowed to equilibrate across a semi- permeable membrane resulting in selective diffusion of unbound test article. The test article concentration is determined in the initial samples and in the filtrates using an appropriate bioanalytical chemistry method (transferred or developed separately) or a microbiological agar diffusion bioassay that uses Bacillus stearothermophilus ATCC 3032 or another suitable species as a test organism.

Reportable Results: Percentage of protein binding.

B0110 Plasma and B0111 Microsome Stability: Plasma and hepatic stability play crucial roles in the ultimate efficacy of therapeutics. Test articles are subject to degradation and modification by a wide range of enzymes in the liver and plasma. To assess stability, the test article is mixed with plasma or liver microsomes from the target host species (nonclinical species or human). At various time points post mixing, aliquots are extracted and analyzed using an appropriate bioanalytical method.

Reportable Results: Half-life and Percent Parent Compound Remaining

www.SouthernResearch.org/Drug-Development Southern Research | in vitro Services B0112 Synergy Testing: Synergy, or combination, testing determines the interaction between two antimicrobials and the resulting impact on the potency of each. The Fractional Inhibitory Concentration (FIC) is determined using the following formula:

FICA + FICB = FIC FICA = MIC of compound A in combination/MIC of compound A alone FICB = MIC of compound B in combination/MIC of compound B alone

The FIC for any given combination is interpreted as follows: Synergy = FIC < 0.5 No interaction = 0.5 < FIC < 4.0 Antagonism = FIC > 4.0

Reportable Results: Fractional Inhibitory Concentration (FIC)

B0113 Minimum Biofilm Inhibition Concentration (MBIC): Bacterial biofilms often demonstrate greater resistance to antibiotics compared with planktonic cells. The Minimum Biofilm Inhibition Concentration (MBIC) assay is a derivative of the MIC wherein the lowest concentration of compound needed to inhibit biofilm formation is determined via a broth microdilution method. Briefly, the challenge bacterium is inoculated into a 96-well tissue culture plate containing appropriate broth and two-fold dilutions of the test compound. The plates are then incubated under conditions that promote biofilm formation. Following growth, total biomass is measured using crystal violet dye. The MBIC is reported as the lowest concentration of drug that completely inhibits biofilm formation.

Reportable Results: Compound concentration (µg/mL)

B0114 Minimum Biofilm Eradication Concentration (MBEC): The MBEC is defined as the lowest concentration of compound needed to eradicate a pre-formed biofilm. In this case, bacterial biofilms are formed on the surface of submerged polystyrene pegs. These pre-formed biofilms are then transferred to a 96-well plate containing broth and two-fold dilutions of the test compounds. After exposure for 18-24 hours at 37°C, the pegs are transferred to fresh media without antibiotic. After an additional incubation period of 8 hours, the optical density of the medium in these wells is measured at 650 nm. The MBEC is reported as the lowest concentration of drug that prevents an increase in optical density of 0.05 or greater.

Reportable Results: Compound concentration (µg/mL)

www.SouthernResearch.org/Drug-Development Southern Research | in vitro Services B0115 Minimum Inhibitory Concentration 90% (MIC 90): Where the classic MIC assay determines the minimum concentration of a test article required to inhibit the growth of a particular isolate, the MIC50 and the MIC90 values represent the minimum concentration of test article that is required to inhibit 50% or 90% of a population of isolates. Typically, at least 30 (and often many more) isolates are tested, representing the full, clinically relevant phenotypic diversity of the target species. Sothern Research maintains MIC50/90 panels for the following bacterial species: Acinetobacter baumannii, Clostridium difficile, Klebsiella pneumonia, Neisseria gonorrhoeae, Pseudomonas aeruginosa, Staphylococcus aureus, and Mycobacterium tuberculosis.

Reportable Results: Compound concentration (µg/mL)

B0116 Opsonophagocytic Killing Assay (OPKA): Typically used as a non-clinical and clinical endpoint for the development of antibacterial vaccines or therapeutic antibodies, the opsonophagocytic killing assay (OPKA) measures the titer of opsonic antibody activity in sera. Briefly, the target bacterium is mixed with whole blood, polymorphonuclear leukocytes (PMNs) or differentiated HL-60 cells and the test sera or antibody. Relative killing is calculated as the percent difference in viable CFU after 0 and 60 minutes of exposure.

Reportable Results: Percent relative killing.

B0117 Opsonophagocytic Uptake Assay (OPUA): Typically used as a non-clinical endpoint for the development of antibacterial vaccines or therapeutic antibodies, the opsonophagocytic uptake assay (OPUA) measures the titer of opsonic antibody activity in sera. Briefly, a fluorescently-labeled target bacterium is mixed with polymorphonuclear leukocytes (PMNs) or differentiated HL-60 cells and the test sera or antibody. Uptake is calculated using flow cytometry.

Reportable Results: Percent uptake.

B0118 Serum Bactericidal Activity (SBA): The serum bactericidal assay (SBA) measures the ability of antibodies to mediate complement- dependent pathogen killing. Log phase bacteria are mixed with serial dilutions of test serum or antibody and an external source of complement. Following incubation, cells are washed and bacterial ATP generation is measured in a microtiter format using a luciferase reporter. Bactericidal serum titers are calculated as the reciprocal serum dilution necessary to obtain a 50% reduction in luminosity compared to control wells.

Reportable Results: Bactericidal titer.

www.SouthernResearch.org/Drug-Development Southern Research | in vivo Services in vivo Testing

B0201 Maximum tolerated dose (MTD): In order to properly interpret data from subsequent efficacy studies, it is necessary to form a basic understanding of the test article toxicology and pharmacokinetics in rodents. The Maximum Tolerated Dose (MTD) is defined as the highest dose of a test article that does not cause overt, acute toxicity. Typically, the MTD is determined using a short duration dose escalation study that include a minimum number of animals.

Reportable Results: Clinical Observations, Morbidity, Mortality, Weight Loss

B0202 Dose Range Finding (DRF): In order to properly interpret data from subsequent efficacy studies, it is necessary to form a basic understanding of the test article toxicology and pharmacokinetics. The Dose Range Finding (DRF) study is used to define the Maximum Repeatable Dose, which is critical for prolonged efficacy models such as chronic sepsis or wound infections. In this study, three or four dose levels of the test article are administered once daily for seven consecutive days. Afterwards, mice are euthanized and gross necropsy and/or histopathological evaluation is used to identify acceptable dosing levels for efficacy testing.

Reportable Results: Clinical Observations, Morbidity, Mortality, Weight Loss, Gross Pathology, Histophathology

B0203 Pharmacokinetic Profiling (PK): Pharmacokinetic (PK) studies are used to determine the rate of test article clearance after administration at the MTD or MRD defined above. This information is critical to proper design and interpretation of subsequent efficacy studies. Serum concentration can be measured using an appropriate bioanalytical assay (e.g., LC-MS/MS, ELISA, qPCR, etc.).

Reportable Results: Serum concentration, PK parameters: Tmax, Cmax, AUC, Vd, CL, T1/2, Kab

B0204 Bioavailability: Bioavailability can be measured during a PK study by adding a second treatment arm and comparing oral or subcutaneous dosing with intravenous dosing.

Reportable Results: Systemically available fraction (plus typical PK results)

www.SouthernResearch.org/Drug-Development Southern Research | in vivo Services B0205 Neutropenic Mouse Thigh (NMTh): The neutropenic mouse thigh is a standard assay used to evaluate the pharmacodynamics characteristics of a test article in vivo. Because viable but antibiotic-damaged bacteria are more susceptible to killing by phagocytic cells, mice are rendered neutropenic for the course of the assay. Mice are challenged with a single inoculation of bacteria into one or both thighs prior to administration of the test article (typically two hours post challenge). At 24-hours post treatment, thighs are aseptically removed, homogenized and plated to determine the total bacterial burden.

Reportable Results: CFU/g and Log10 CFU reduction from control group

B0206 Neutropenic Mouse Lung (NMLu): Because the pharmacokinetics of the lung compartment often differs from that of the serum and muscle, the neutropenic mouse lung assay is used to evaluate the pharmacodynamics of a test article during infection of the lung. Mice are rendered neutropenic for the course of the assay and are challenged with a single intranasal inoculation of bacteria. Two hours after challenge, the test article is administered. At 24-hours post treatment, lungs are aseptically removed, homogenized and plated to determine the total bacterial burden.

Reportable Results: CFU/g tissue and Log10 CFU reduction from control group

B0207 Acute Murine Sepsis/Peritonitis: The acute sepsis/peritonitis model is an ideal platform to screen for initial in vivo efficacy relative to systemic indications in an immune competent host animal. A single, high-dose challenge of bacteria is used to generate acute bacteremia and endotoxemia. Antibacterial and control agents are administered following challenge and blood is harvested from all animals at 12 hours post treatment. Symptoms are recorded on a standardized scoresheet at 6 hours and 12 hours post-treatment and every 12 hours thereafter until the end of the study. Bacterial burden in the spleen or kidney of surviving animals is analyzed at 48-72 hours post-treatment.

Reportable Results: Survival, Clinical Score, CFU/ml blood, CFU/g tissue, Log10 CFU reduction from control group

B0208 Chronic Murine Sepsis (Cecal Ligation and Puncture): Surgical ligation and puncture of the cecum induces a prolonged, low-level, polymicrobial septicemia. This procedure generally leads to 80-90% mortality in untreated mice by day 5. All mice are observed twice daily for mortality and surviving mice are weighed and scored for severity of disease symptoms using a standardized score sheet.

Reportable Results: Survival, Clinical Score, Body Weight

www.SouthernResearch.org/Drug-Development Southern Research | in vivo Services B0209 Murine Acute Pneumonia: The acute pneumonia model is an ideal platform to screen for initial in vivo efficacy relative to lung indications in an immune competent host. C57Bl/6 mice are challenged with a single dose of bacteria delivered intranasally. Test and control articles are administered two hours after challenge. At 6- and 24- hours after treatment, mice are euthanized and the bacterial burden in the blood and lungs are determined by serial dilution.

Reportable Results: CFU/ml blood, CFU/g tissue, Log10 CFU reduction from control group

B0210 Skin Infection – Tape-stripping: The skin tape stripping model measures the efficacy of a test article in a setting of superficial skin infection where damage of underlying layers of the skin is undesirable or irrelevant for the desired indication. In this model, superficial damage is achieved by successive application of an adhesive bandage to strip off the fur and epidermis. After stripping, bacteria are allowed to adhere to the wound for 4 hours prior to initiation of treatment. All mice are observed at least twice daily for mortality and surviving mice are weighed and scored for wound severity. At the end of the experiment, the wound area is excised, homogenized, and bacterial burden is determined by serial dilution.

Reportable Results: Wound Severity, CFU/g tissue, Log10 CFU reduction from control group

B0211 Surgical Site Infection—Suture: The suture model measures efficacy of a test article in the setting of a surgical sight infection. A surgical suture is contaminated with the bacterium of interest prior to implantation into an incision wound on the back of mice. The wound is closed with a single, non-contaminated suture and test or control articles treatment is initiated 2 hours post-surgery. Mice are euthanized at 24 hours post-treatment and the wound area is excised, homogenized and plated to determine the bacterial burden.

Reportable Results: CFU/g tissue, Log10 CFU reduction from control group

B0212 Wound Infection—Punch Biopsy: The punch biopsy wound model measures the efficacy of a test article in the setting of a full-thickness skin wound. A large skin biopsy punch is used to great a 6.0 mm wound on the back directly above the thoracic spinal column and adjacent musculature. Bacteria are allowed to absorb to the wound prior to covering with adherent dressing. Treatment is initiated 4 hours after wounding. Mice are euthanized after 7 days, the wound is excised, homogenized and plated to determine the bacterial burden.

Reportable Results: CFU/g tissue, Log10 CFU reduction from control group

www.SouthernResearch.org/Drug-Development Southern Research | in vivo Services B0213 Transcervical Chlamydia Persistence: In order to meet the needs of Chlamydia vaccine developers, Southern Research developed a C57BL/6 model of infection/colonization by human Chlamydia trachomatis isolates. Infectious elementary bodies are delivered directly to the upper reproductive tract using a non-surgical technique that results in infection/colonization for several days. The assay has been used successfully with serovars D and L2.

Reportable Results: Bacterial burden in the upper genital tract (by qPCR)

B0214 Pharmacodynamic Dose Fractionation: PK-PD based non-clinical data helps de-risk drug development and strengthens NDA submissions because it is critical to setting a successful dosing regimen. A dose fractionation strategy, whereby the same total accumulative dose is given across different numbers of administrations, is used to identify both the PK-PD index associated with efficacy and the index magnitude required for efficacy.

Reportable Results: PK-PD indices versus Log10 CFU bacteria at the site of infection.

B0215 in vivo Resistance Monitoring: Often performed in conjunction with B0214 Pharmacodynamic dose fractionation, this assay measures the frequency of resistant mutant generation following different dosing regimens. Bacteria are harvested from target organs at specific time points and plated on media containing the test article.

Reportable Results: Frequency of resistance emergence

B0216 in vivo Post-Antibiotic Effect (PAE): PK-PD based non-clinical data helps de-risk drug development and strengthens NDA submissions because it is critical to setting a successful dosing regimen. The post antibiotic effect (PAE) is defined as persistent suppression of bacterial growth following exposure. Analysis of the in vivo PAE helps inform dosing strategy and, in general, antibiotics that induce a stronger PAE are more desirable because they would require less frequent administration. This analysis is typically performed using the neutropenic mouse thigh assay. The in vivo PAE is calculated using the formula PAE = T – C – M where M is the time serum levels exceed the MIC, T is the time required for CFU in the thighs of treated mice to increase by 1

Log10 above the count at time M, and C is the time required for CFU in the thighs of untreated mice to increase by 1 Log10.

Reportable Results: PAE (hours)

www.SouthernResearch.org/Drug-Development Southern Research | in vivo Services B0217 Vaccine Immunogenicity Evaluation: Proof-of-concept immunogenicity studies are critical to moving a candidate vaccine from early development into the clinic. Data from these experiments is used to establish the immunological characteristics of the product and may guide selection of doses, schedules and routes of administration for subsequent efficacy studies in nonclinical models and in clinical trials. Southern Research can support immunogenicity testing in a number of nonclinical species including rodents and non-human primates. In most cases, both antigen binding and functional activity (e.g., neutralization or opsonophagocytic activity) can be measured.

Reportable Results: Total and functional antibody titer over time.

IND-Enabling Toxicology: For nearly 50 years, Southern Research has conducted nonclinical safety assessments for a wide variety of chemical entities—from small molecule pharmaceuticals to biologics to vaccines. Today, we continue to focus on the unique needs of each client in order to execute the most appropriately designed studies for each situation, thus ensuring that the path to regulatory approval is as smooth as possible. Our toxicologists are backed by highly trained technical staff and an array of in-house core services including on-site bioanalytical, histopathology and clinical pathology support as well as robust, state-of-the-art systems for data capture and management. Study types include acute and repeat dose (subchronic and chronic) toxicity, carcinogenicity/oncogenicity, developmental and reproductive toxicology (DART), neurotoxicity and neurobehavioral assessment.

www.SouthernResearch.org/Drug-Development Southern Research | Clinical Trial Support Services

Clinical Trial Endpoint Analysis

B0301 Opsonophagocytic Killing Assay (OPKA): Typically used as a non-clinical and clinical endpoint for the development of antibacterial vaccines or therapeutic antibodies, the opsonophagocytic killing assay (OPKA) measures the titer of opsonic antibody activity in sera. Briefly, the target bacterium is mixed with polymorphonuclear leukocytes (PMNs) or differentiated HL-60 cells and the test sera. Relative killing is calculated as the percent difference between viable CFU after 0 and 60 minutes of exposure.

Reportable Results: Percent relative killing.

B0302 Opsonophagocytic Uptake Assay (OPUA): The opsonophagocytic uptake assay (OPUA) measures the titer of opsonic antibody activity in sera. Briefly, a fluorescently-labeled target bacterium is mixed with polymorphonuclear leukocytes (PMNs) or differentiated HL-60 cells and the test sera. Uptake is calculated using flow cytometry.

Reportable Results: Percent uptake.

B0303 Serum Bactericidal Activity (SBA): The serum bactericidal assay (SBA) measures the ability of antibodies present in serum to mediate complement-dependent pathogen killing. Log phase bacteria are mixed with serial dilutions of test sera and an external source of complement. Following incubation, cells are washed and bacterial ATP generation is measured in a microtiter format using a luciferase reporter. Bactericidal serum titers are calculated as the reciprocal serum dilution necessary to obtain a 50% reduction in luminosity compared to control wells.

Reportable Results: Bactericidal serum titer.

Additional Bioanalytical and Immunological Endpoint Assays: Southern Research is a highly collaborative, full-service Contract Research Organization. For nearly 50 years, we have stayed at the forefront of biomedical research and drug development. This perspective allows our scientists to apply their vast experiences and deep expertise in service of your specific needs. We can develop fit-for-purpose bioanalytical and immunological assays from early discovery phase to clinical trials including Anti-drug Antibody (ADA) Assays, Neutralizing Antibody (Nab) quantification, and Serum Drug Concentration.

www.SouthernResearch.org/Drug-Development MIC/MBC Panels

Drug ResistantMIC90 Strains panelESKAPE PanelMDR PanelCRE Panel ESBL-PanelRespiratoryCF panel Panel STI Panel Bacterial MeningitisEndocarditis PanelGastroenteritis Panel Wound Panel Infection Panel Achromobacter xyloxidans x x Bacillus cereus Acinetobacter baumannii x x x x x x x Aggregatibacter sp. x Atopobium vaginae x Burkholderia cepacia complex x x x x Burkholderia multivorans x Campylobacter jejuni x Cardiobacterium hominis x Chlamydia trachomatis x Clostridium difficile x x x Eikenella corrodens x Enteric Group 137 x Enterobacter aerogenes x Enterobacter cloacae x x x x Enterococcus faecium x x Escherichia coli x x x x x x Gardnerella vaginalis x Haemophilus influenzae x x x Haemophilus sp. x Helicobacter pylori x Kingella sp. x Klebsiella pneumoniae x x x x x x Listeria monocytogenes x Mobiluncus curtisii x Mobiluncus mulieris x Mycobacterium abscessus x Mycoplasma genitalium x Neisseria gonorrhoeae x x x Neisseria meningitidis x Proteus mirabilis x x Pseudomonas aeruginosa x x x x x x x Salmonella enterica x Shigella dysenteriae x Staphylococcus aureus x x x x x x x x x Staphylococcus epidermidis x x Stenotrophomonas maltophilia x x x Streptococcus agalactiae x Streptococcus gordonii x Streptococcus oralis x Streptococcus pneumonia x x x x Streptococcus pyogenes x Viridians streptococci x Yersinia enterocolitica x Additional strains available

Drug ResistantMIC90 Strains panel Achromobacter xyloxidans x x Bacillus cereus Chlamydophila pneumoniae Citrobacter freundii x Citrobacter koseri Clostridium perfringens Enterococcus avium x Enterococcus faecalis x Klebsiella oxytoca x Lactobacillus acidophilus Legionella pneumoniae Listeria monocytogenes Moraxella catarrhalis Morganella morganii Mycobacterium avium Mycobacterium intracellularae Mycobacterium smegmatis Mycobacterium tuberculosis x x Mycobacterium ulcerans Mycoplasma pneumoniae Propionibacterium acnes Proteus vulgaris x Providencia stuartii Salmonella enterica x Serratia marcescens x Shewanella putrefaciens x Vibrio cholera